Therapeutically active compounds and their methods of use

ABSTRACT

Provided are methods of treating a cancer characterized by the presence of a mutant allele of IDH1/2 comprising administering to a subject in need thereof a compound described here.

CLAIM OF PRIORITY

This application is a continuation of U.S. Ser. No. 15/809,325, filedNov. 10, 2017, which is a continuation of U.S. Ser. No. 15/279,146,filed Sep. 28, 2016, which is a continuation of U.S. Ser. No.13/745,005, filed Jan. 18, 2013, which claims priority under 35 U.S.C. §119 from International Application No. PCT/CN2012/070601, filed Jan. 19,2012, each of which is incorporated herein by reference in its entirety.

BACKGROUND OF INVENTION

Isocitrate dehydrogenases (IDHs) catalyze the oxidative decarboxylationof isocitrate to 2-oxoglutarate (i.e., α-ketoglutarate). These enzymesbelong to two distinct subclasses, one of which utilizes NAD(+) as theelectron acceptor and the other NADP(+). Five isocitrate dehydrogenaseshave been reported: three NAD(+)-dependent isocitrate dehydrogenases,which localize to the mitochondrial matrix, and two NADP(+)-dependentisocitrate dehydrogenases, one of which is mitochondrial and the otherpredominantly cytosolic. Each NADP(+)-dependent isozyme is a homodimer.

IDH1 (isocitrate dehydrogenase 1 (NADP+), cytosolic) is also known asIDH; IDP; IDCD; IDPC or PICD. The protein encoded by this gene is theNADP(+)-dependent isocitrate dehydrogenase found in the cytoplasm andperoxisomes. It contains the PTS-1 peroxisomal targeting signalsequence. The presence of this enzyme in peroxisomes suggests roles inthe regeneration of NADPH for intraperoxisomal reductions, such as theconversion of 2,4-dienoyl-CoAs to 3-enoyl-CoAs, as well as inperoxisomal reactions that consume 2-oxoglutarate, namely thealpha-hydroxylation of phytanic acid. The cytoplasmic enzyme serves asignificant role in cytoplasmic NADPH production.

The human IDH1 gene encodes a protein of 414 amino acids. The nucleotideand amino acid sequences for human IDH1 can be found as GenBank entriesNM_005896.2 and NP_005887.2 respectively. The nucleotide and amino acidsequences for IDH1 are also described in, e.g., Nekrutenko et al., Mol.Biol. Evol. 15:1674-1684(1998); Geisbrecht et al., J. Biol. Chem.274:30527-30533(1999); Wiemann et al., Genome Res. 11:422-435(2001); TheMGC Project Team, Genome Res. 14:2121-2127(2004); Lubec et al.,Submitted (DEC-2008) to UniProtKB; Kullmann et al., Submitted (JUN-1996)to the EMBL/GenBank/DDBJ databases; and Sjoeblom et al., Science314:268-274(2006).

Non-mutant, e.g., wild type, IDH1 catalyzes the oxidativedecarboxylation of isocitrate to α-ketoglutarate thereby reducing NAD⁺(NADP⁺) to NADH (NADPH), e.g., in the forward reaction:Isocitrate+NAD⁺ (NADP⁺)→α-KG+CO₂+NADH (NADPH)+H⁺.

It has been discovered that mutations of IDH1 present in certain cancercells result in a new ability of the enzyme to catalyze theNAPH-dependent reduction of α-ketoglutarate to R(−)-2-hydroxyglutarate(2HG). The production of 2HG is believed to contribute to the formationand progression of cancer (Dang, L et al, Nature 2009, 462:739-44).

The inhibition of mutant IDH1 and its neoactivity is therefore apotential therapeutic treatment for cancer. Accordingly, there is anongoing need for inhibitors of IDH1 mutants having alpha hydroxylneoactivity.

SUMMARY OF INVENTION

Described herein are methods of treating a cancer characterized by thepresence of a mutant allele of IDH1 or IDH2. The methods comprise thestep of administering to a subject in need thereof a compound of formulaI, or a pharmaceutically acceptable salt, tautomer, isotopologue orhydrate thereof, wherein:

R¹ is optionally substituted C₄-C₆ carbocyclyl;

each R² and R³ is independently selected from optionally substitutedaryl or optionally substituted heteroaryl;

R⁴ is alkyl, optionally substituted aryl, optionally substitutedheteroaryl, optionally substituted aralkyl, or optionally substitutedheteroaralkyl;

ring A is 4-6 membered non-aromatic ring having 0-1 additionalheteroatoms selected from N, O or S, wherein ring A is optionallysubstituted with one or two R⁵ groups;

each R⁵ is independently halo; —CF₃; —CN; —OR⁶; —N(R⁶)₂; —C(O)C₁-C₄alkyl; C₁-C₄ haloalkyl; C₁-C₄ alkyl optionally substituted with —OR⁶ or—N(R⁶)₂; —O—C₁-C₄ alkyl optionally substituted with halo, —OR⁶ or—N(R⁶)₂; —SO₂N(R⁶)₂; —SO₂(C₁-C₄ alkyl); —NR⁶SO₂R⁶; C₃-C₅ carbocyclyloptionally substituted with one or two R⁶ groups; —O—(C₃-C₆ carbocyclyloptionally substituted with one or two R⁶ groups); 5-6 memberedheteroaryl; —C₁-C₄ alkyl-C(O)O—C₁-C₄ alkyl; or —C(O)O—C₁-C₄ alkyl; or

each R⁶ is independently H or C₁-C₃ alkyl.

The compound of formula I inhibits mutant IDH1/2, particularly mutantIDH1 having alpha hydroxyl neoactivity. Also described herein arepharmaceutical compositions comprising a compound of formula I.

DETAILED DESCRIPTION OF THE INVENTION

The details of construction and the arrangement of components set forthin the following description or illustrated in the drawings are notmeant to be limiting. Other embodiments and different ways to practicethe invention are expressly included. Also, the phraseology andterminology used herein is for the purpose of description and should notbe regarded as limiting. The use of “including,” “comprising,” or“having,” “containing”, “involving”, and variations thereof herein, ismeant to encompass the items listed thereafter and equivalents thereofas well as additional items.

Definitions

The term “halo” or “halogen” refers to any radical of fluorine,chlorine, bromine or iodine.

The term “alkyl” refers to a hydrocarbon chain that may be a straightchain or branched chain, containing the indicated number of carbonatoms. For example, C₁-C₁₂ alkyl indicates that the group may have from1 to 12 (inclusive) carbon atoms in it. The term “haloalkyl” refers toan alkyl in which one or more hydrogen atoms are replaced by halo, andincludes alkyl moieties in which all hydrogens have been replaced byhalo (e.g., perfluoroalkyl). The terms “arylalkyl” or “aralkyl” refer toan alkyl moiety in which an alkyl hydrogen atom is replaced by an arylgroup. Arylalkyl or aralkyl includes groups in which more than onehydrogen atom has been replaced by an aryl group. Examples of“arylalkyl” or “aralkyl” include benzyl, 2-phenylethyl, 3-phenylpropyl,9-fluorenyl, benzhydryl, and trityl groups. The terms “heteroarylalkyl”or “heteroaralkyl” refer to an alkyl moiety in which an alkyl hydrogenatom is replaced by a heteroaryl group. Heteroarylalkyl or heteroaralkylincludes groups in which more than one hydrogen atom has been replacedby a heteroaryl group.

The term “alkylene” refers to a divalent alkyl, e.g., —CH₂—, —CH₂CH₂—,and —CH₂CH₂CH₂—.

The term “alkenyl” refers to a straight or branched hydrocarbon chaincontaining 2-12 carbon atoms and having one or more double bonds.Examples of alkenyl groups include, but are not limited to, allyl,propenyl, 2-butenyl, 3-hexenyl and 3-octenyl groups. One of the doublebond carbons may optionally be the point of attachment of the alkenylsubstituent. The term “alkynyl” refers to a straight or branchedhydrocarbon chain containing 2-12 carbon atoms and characterized inhaving one or more triple bonds. Examples of alkynyl groups include, butare not limited to, ethynyl, propargyl, and 3-hexynyl. One of the triplebond carbons may optionally be the point of attachment of the alkynylsubstituent.

The term “alkoxy” refers to an —O-alkyl radical. The term “haloalkoxy”refers to an alkoxy in which one or more hydrogen atoms are replaced byhalo, and includes alkoxy moieties in which all hydrogens have beenreplaced by halo (e.g., perfluoroalkoxy).

The term “carbocyclyl” refers to a monocyclic, bicyclic or tricyclic,hydrocarbon ring system that is not fully aromatic, wherein any ringatom capable of substitution can be substituted by one or moresubstituents. A carbocyclyl can be fully or partially saturated. Abicyclic or tricylic carbocyclyl may contain one (in the case of abicycle) or up to two (in the case of a tricycle) aromatic rings, aslong as at least one ring in the carbocyclyl is non-aromatic. Unlessotherwise specified, any ring atom capable of substitution in acarbocyclyl can be substituted by one or more substituents.

The term “aryl” refers to a fully aromatic monocyclic, bicyclic, ortricyclic hydrocarbon ring system. Examples of aryl moieties are phenyl,naphthyl, and anthracenyl. Unless otherwise specified, any ring atom inan aryl can be substituted by one or more substituents.

The term “cycloalkyl” as employed herein refers to a saturated cyclic,bicyclic, tricyclic, or polycyclic hydrocarbon group. Unless otherwisespecified, any ring atom can be substituted by one or more substituents.The cycloalkyl groups can contain fused rings. Fused rings are ringsthat share a common carbon atom. Examples of cycloalkyl moietiesinclude, but are not limited to, cyclopropyl, cyclohexyl,methylcyclohexyl, adamantyl, and norbornyl. Unless otherwise specified,any ring atom can be substituted by one or more substituents.

The term “heterocyclyl” refers to a monocyclic, bicyclic or tricyclic,ring structure that is not fully aromatic and includes one to fourheteroatoms independently selected from N, O, or S in one or more of therings. A heterocyclyl can be fully or partially saturated. A bicyclic ortricylic heterocyclyl may contain one (in the case of a bicycle) or upto two (in the case of a tricycle) aromatic rings, as long as at leastone ring in the heterocyclyl is non-aromatic. Unless otherwisespecified, any ring atom capable of substitution in a heterocyclyl canbe substituted by one or more substituents. Heterocyclyl groups include,for example, thiophene, thianthrene, furan, pyran, isobenzofuran,chromene, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole,isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine,indolizine, isoindole, indole, indazole, purine, quinolizine,isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline,quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine,acridine, pyrimidine, phenanthroline, phenazine, phenarsazine,phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane,oxazole, piperidine, piperazine, morpholine, lactones, lactams such asazetidinones and pyrrolidinones, sultams, sultones, and the like.

The term “heteroaryl” refers to a monocyclic, bicyclic, or tricyclicring system having 1-3 heteroatoms if monocyclic, 1-6 heteroatoms ifbicyclic, or 1-9 heteroatoms if tricyclic, said heteroatomsindependently selected from O, N, or S, wherein each ring in aheteroaryl is fully aromatic. Unless otherwise specified, any ring atomcapable of substitution in a heteroaryl can be substituted by one ormore substituents. The terms “hetaralkyl” and “heteroaralkyl”, as usedherein, refers to an alkyl group substituted with a heteroaryl group.The ring heteroatoms of the compounds provided herein include N—O, S(O),and S(O)₂.

The term “substituted” refers to the replacement of a hydrogen atom withanother moiety. Typical substituents include alkyl (e.g., C1, C2, C3,C4, C5, C6, C7, C8, C9, C10, C11, C12 straight or branched chain alkyl),cycloalkyl, haloalkyl (e.g., perfluoroalkyl such as CF₃), aryl,heteroaryl, aralkyl, heteroaralkyl, heterocyclyl, alkenyl, alkynyl,cycloalkenyl, heterocycloalkenyl, alkoxy, haloalkoxy (e.g.,perfluoroalkoxy such as OCF₃), halo, hydroxy, carboxy, carboxylate,cyano, nitro, amino, alkyl amino, SO₃H, sulfate, phosphate,methylenedioxy (—O—CH₂—O— wherein oxygens are attached to vicinalatoms), ethylenedioxy, oxo (not a substituent on heteroaryl), thioxo(e.g., C═S) (not a substituent on heteroaryl), imino (alkyl, aryl,aralkyl), S(O)_(n)alkyl (where n is 0-2), S(O)_(n) aryl (where n is0-2), S(O)_(n) heteroaryl (where n is 0-2), S(O)_(n) heterocyclyl (wheren is 0-2), amine (mono-, di-, alkyl, cycloalkyl, aralkyl, heteroaralkyl,aryl, heteroaryl, and combinations thereof), ester (alkyl, aralkyl,heteroaralkyl, aryl, heteroaryl), amide (mono-, di-, alkyl, aralkyl,heteroaralkyl, aryl, heteroaryl, and combinations thereof), sulfonamide(mono-, di-, alkyl, aralkyl, heteroaralkyl, and combinations thereof).In one aspect, the substituents on a group are independently any onesingle, or any subset of the aforementioned substituents. In anotheraspect, a substituent may itself be substituted with any one of theabove substituents.

The term “tautomer” refers to each of two or more isomers of a compound(e.g., a compound described herein) that exist together in equilibrium,and are readily interchangeable by migration of a hydrogen atom orproton, accompanied by a switch of a single bond and an adjacent doublebond.

As used herein, the term “elevated levels of 2HG” means 10%, 20% 30%,50%, 75%, 100%, 200%, 500% or more 2HG than is present in a subject thatdoes not carry a mutant IDH1 or IDH2 allele. The term “elevated levelsof 2HG” may refer to the amount of 2HG within a cell, within a tumor,within an organ comprising a tumor, or within a bodily fluid.

The term “bodily fluid” includes one or more of amniotic fluidsurrounding a fetus, aqueous humour, blood (e.g., blood plasma), serum,Cerebrospinal fluid, cerumen, chyme, Cowper's fluid, female ejaculate,interstitial fluid, lymph, breast milk, mucus (e.g., nasal drainage orphlegm), pleural fluid, pus, saliva, sebum, semen, serum, sweat, tears,urine, vaginal secretion, or vomit.

As used herein, the terms “inhibit” or “prevent” include both completeand partial inhibition and prevention. An inhibitor may completely orpartially inhibit.

The term “treat” means decrease, suppress, attenuate, diminish, arrest,or stabilize the development or progression of a cancer (e.g., a cancerdelineated herein), lessen the severity of the cancer or improve thesymptoms associated with the cancer.

As used herein, an amount of a compound effective to treat a disorder,or a “therapeutically effective amount” refers to an amount of thecompound which is effective, upon single or multiple dose administrationto a subject, in treating a cell, or in curing, alleviating, relievingor improving a subject with a disorder beyond that expected in theabsence of such treatment.

As used herein, the term “subject” is intended to include human andnon-human animals. Exemplary human subjects include a human patienthaving a disorder, e.g., a disorder described herein or a normalsubject. The term “non-human animals” of the invention includes allvertebrates, e.g., non-mammals (such as chickens, amphibians, reptiles)and mammals, such as non-human primates, domesticated and/oragriculturally useful animals, e.g., sheep, dog, cat, cow, pig, etc.

Compounds

Provided is a compound having formula I or a pharmaceutically acceptablesalt, tautomer, isotopologue or hydrate thereof, wherein:

R¹ is optionally substituted C₄-C₆ carbocyclyl;

each R² and R³ is independently selected from optionally substitutedaryl or optionally substituted heteroaryl;

R⁴ is alkyl, optionally substituted aryl, optionally substitutedheteroaryl, optionally substituted aralkyl, or optionally substitutedheteroaralkyl;

ring A is 4-6 membered non-aromatic ring having 0-1 additionalheteroatoms selected from N, O or S, wherein ring A is optionallysubstituted with one or two R⁵ groups;

each R⁵ is independently halo; —CF₃; —CN; —OR⁶; —N(R⁶)₂; —C(O)C₁-C₄alkyl; C₁-C₄ haloalkyl; C₁-C₄ alkyl optionally substituted with —OR⁶ or—N(R⁶)₂; —O—C₁-C₄ alkyl optionally substituted with halo, —OR⁶ or—N(R⁶)₂; —SO₂N(R⁶)₂; —SO₂(C₁-C₄ alkyl); —NR⁶SO₂R⁶; C₃-C₅ carbocyclyloptionally substituted with one or two R⁶ groups; —O—(C₃-C₆ carbocyclyloptionally substituted with one or two R⁶ groups); 5-6 memberedheteroaryl; —C₁-C₄ alkyl-C(O)O—C₁-C₄ alkyl; or —C(O)O—C₁-C₄ alkyl; or

each R⁶ is independently H or C₁-C₃ alkyl.

Provided is also a compound having formula I or a pharmaceuticallyacceptable salt or hydrate thereof, wherein:

R¹ is optionally substituted C₄-C₆ carbocyclyl;

each R² and R³ is independently selected from optionally substitutedaryl or optionally substituted heteroaryl;

R⁴ is alkyl, optionally substituted aryl, optionally substitutedheteroaryl, optionally substituted aralkyl, or optionally substitutedheteroaralkyl;

ring A is 4-6 membered non-aromatic ring having 0-1 additionalheteroatoms selected from N, O or S, wherein ring A is optionallysubstituted with one or two R⁵ groups;

each R⁵ is independently halo, —CF₃, —CN, —OR⁶, —N(R⁶)₂, —C(O)CH₃; C₁-C₃haloalkyl, C₁-C₃ alkyl optionally substituted with —OR⁶ or —N(R⁶)₂; or

each R⁶ is independently H or C₁-C₃ alkyl.

Provided is also a compound having formula I or a pharmaceuticallyacceptable salt, tautomer, isotopologue or hydrate thereof, wherein:

R¹ is C₄-C₆ carbocyclyl optionally substituted with one to three R⁷groups;

each R² and R³ is independently selected from aryl or heteroaryl,wherein said aryl or heteroaryl is independently optionally substitutedwith one to three R⁷ groups;

R⁴ is alkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl, wherein saidaryl, heteroaryl, aralkyl, and heteroaralkyl are each independentlyoptionally substituted with one to three R⁷ groups;

ring A is 4-6 membered non-aromatic ring having 0-1 additionalheteroatoms selected from N, O or S, wherein ring A is optionallysubstituted with one or two R⁵ groups;

each R⁵ and R⁷ is independently halo; —CF₃; —CN; —OR⁶; —N(R⁶)₂;—C(O)C₁-C₄ alkyl; C₁-C₄ haloalkyl; C₁-C₄ alkyl optionally substitutedwith —OR⁶ or —N(R⁶)₂; —O—C₁-C₄ alkyl optionally substituted with halo,—OR⁶ or —N(R⁶)₂; —SO₂N(R⁶)₂; —SO₂(C₁-C₄ alkyl); —S(O)—C₁₋₄ alkyl,—NR⁶SO₂R⁶; C₃-C₅ carbocyclyl optionally substituted with one or two R⁶groups; —O—(C₃-C₆ carbocyclyl optionally substituted with one or two R⁶groups); 5-6 membered heteroaryl; —C₁-C₄ alkyl-C(O)O—C₁-C₄ alkyl; or—C(O)O—C₁-C₄ alkyl; or

each R⁶ is independently H or C₁-C₄ alkyl.

Provided is also a compound having formula I or a pharmaceuticallyacceptable salt, tautomer, isotopologue or hydrate thereof, wherein:

R¹ is C₄-C₆ carbocyclyl optionally substituted with one to three R⁷groups;

each R² and R³ is independently selected from aryl or heteroaryl,wherein said aryl or heteroaryl is independently optionally substitutedwith one to three R⁷ groups;

R⁴ is alkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl, wherein saidaryl, heteroaryl, aralkyl, and heteroaralkyl are each independentlyoptionally substituted with one to three R⁷ groups;

ring A is 4-6 membered non-aromatic ring having 0-1 additionalheteroatoms selected from N, O or S, wherein ring A is optionallysubstituted with one or two R⁵ groups;

each R⁵ and R⁷ is independently halo, —CF₃, —CN, —OR⁶, —N(R⁶)₂,—C(O)CH₃; C₁-C₃ haloalkyl, C₁-C₃ alkyl optionally substituted with OR⁶or —N(R⁶)₂; or

each R⁶ is independently H or C₁-C₃ alkyl.

In one embodiment, R¹ is optionally substituted C₄-C₆ cycloalkyl. In oneaspect of this embodiment, R¹ is C₄-C₆ cycloalkyl optionally substitutedwith one to three R⁷ groups. In another aspect of this embodiment, R¹ isC₄, C₅, or C₆ cycloalkyl optionally substituted with one to two R⁷groups and R⁷ is halo. In another aspect of this embodiment, R¹ is C₄ orC₆ cycloalkyl optionally substituted with one to two R⁷ groups and R⁷ ishalo. In yet another aspect of this embodiment, R¹ is

In yet another aspect of this embodiment, R¹ is

In another embodiment, R² is optionally substituted aryl. In one aspectof this embodiment, R² is aryl optionally substituted with one to threeR⁷ groups. In another aspect of this embodiment, R² is phenyl optionallysubstituted with one to two R⁷ groups and R⁷ is —Cl.

In another embodiment, R³ is optionally substituted aryl or optionallysubstituted aryl heteroaryl. In one aspect of this embodiment, R³ isoptionally substituted heteroaryl. In another aspect of this embodiment,R³ is heteroaryl optionally substituted with one to three R⁷ group. Inyet another aspect of this embodiment, R³ is pyridinyl, indazolyl,benzoimidazolyl, indolyl, or N-methylindolyl, wherein each R³ isoptionally substituted with one R⁷ wherein R⁷ is F. In another aspect ofthis embodiment, R³ is optionally substituted aryl. In another aspect ofthis embodiment, R³ is aryl optionally substituted with one to three R⁷groups. In yet another aspect of this embodiment, R³ is phenyloptionally substituted with one R⁷ wherein R⁷ is —F. In yet anotheraspect of this embodiment, R³ is phenyl optionally substituted with oneor two R⁷s wherein each R⁷ is independently halo; —CN; —N(R⁶)₂; C₁-C₄alkyl optionally substituted with —OR⁶; —O—C₁-C₄ alkyl optionallysubstituted with halo, or —OR⁶; —SO₂N(R⁶)₂; —SO₂(C₁-C₄ alkyl);—S(O)—C₁₋₄ alkyl, —NR⁶SO₂R⁶; C₃-C₅ carbocyclyl optionally substitutedwith one R⁶; —O—(C₃-C₆ carbocyclyl); 5-membered heteroaryl. In yetanother aspect of this embodiment, R³ is phenyl optionally substitutedwith one or two les wherein each R⁷ is independently —F, —SO₂NH₂,—SO₂CH₃, —S(O)CH₃, —CN, methoxy, —OCH₂OH, —CH₂OH, —SO₂N(CH₃)₂,—SO₂NHCH₃, —NHSO₂CH₃, —CH₂CH₂OH, —N(CH₃)₂, t-butyl, cyclopropyl,—C(OH)(CH₃)₂, —OCF₃, —OCHF₂, —O-cyclopropyl, 1-methyl-cyclopropyl, orpyrazolyl.

In another embodiment, R⁴ is optionally substituted aryl, optionallysubstituted heteroaryl, optionally substituted aralkyl, or optionallysubstituted heteroaralkyl. In one aspect of this embodiment, R⁴ is aryl,heteroaryl, aralkyl, or heteroaralkyl, wherein said aryl, heteroaryl,aralkyl, and heteroaralkyl are each independently optionally substitutedwith one to three R⁷ groups. In another aspect of this embodiment, R⁴ isaryl or heteroaryl, each aryl or heteroaryl is optionally substitutedwith one to three R⁷ groups. In another aspect of this embodiment, R⁴ is6-membered aryl or 5-6 membered heteroaryl, wherein said aryl orheteroaryl is optionally substituted with one to three R⁷ groups. In yetanother aspect of this embodiment, R⁴ is:

wherein each member of R⁴ is optionally substituted with one or two R⁷groups and each R⁷ is independently F, Cl, methyl, CF₃, CN, OMe, orN(R⁶)₂. In yet another aspect of this embodiment, R⁴ is:

wherein each R¹⁰⁰ is independently H, methyl, F, Cl, CF₃, CN, OCH₃, orN(R⁶)₂. In yet another aspect of this embodiment, R⁴ is:

wherein R¹⁰⁰ is H, methyl, F, Cl, CF₃, CN, OCH₃, or N(R⁶)₂. In yetanother embodiment, R⁴ is:

wherein R¹⁰⁰ is H, methyl, Cl, CF₃, CN, OCH₃, or N(R⁶)₂ and R¹⁰¹ is H, For methyl.

In another embodiment, ring A is

wherein

denotes ring A's attachment to the amide moiety of formula and

denotes ring A's attachment to R⁴; and each member of ring A isoptionally substituted with one or two R⁵ groups. In another embodiment,ring A is

wherein

denotes ring A's attachment to the amide moiety of formula and

denotes ring A's attachment to R⁴; and each member of ring A isoptionally substituted with one or two R⁵ groups. In one aspect of thisembodiment, each R⁵ is independently halo; —OR⁶; —C(O)C₁-C₄ alkyl; C₁-C₄alkyl optionally substituted with —OR⁶; —C₃-C₅ carbocyclyl optionallysubstituted with one or two R⁶ groups; —C₁-C₄ alkyl-C(O)O—C₁-C₄ alkyl;or —C(O)O—C₁-C₄ alkyl. In one aspect of this embodiment, each R⁵ isindependently —OH, —F, —CH₂CH₂OH, —CH₂C(O)OCH₂CH₃, —C(O)O-t-butyl,cyclopropyl, methyl or —C(O)CH₃. In one aspect of this embodiment, eachR⁵ is independently methyl or —C(O)CH₃. In another aspect of thisembodiment, ring A is:

In another aspect of this embodiment, ring A is:

Provided is also a compound having formula II or a pharmaceuticallyacceptable salt or hydrate thereof, wherein R¹, R², R³, ring A and R⁴are as defined in formula I or any one of the above embodiments.

Provided is also a compound having formula II-a or a pharmaceuticallyacceptable salt or hydrate thereof, wherein R¹, R⁴, ring A and R⁷ are asdefined in formula I or any one of the above embodiments.

Provided is also a compound having formula II-a-1 or a pharmaceuticallyacceptable salt or hydrate thereof, wherein R¹, R⁴, ring A and R⁷ are asdefined in formula I or any one of the above embodiments and R¹⁰ is CR¹¹or N wherein R¹¹ is —F, —SO₂NH₂, —SO₂CH₃, —CN, methoxy, —OCH₂OH, —CH₂OH,—SO₂N(CH₃)₂, —SO₂NHCH₃, —NHSO₂CH₃, —CH₂CH₂OH, —N(CH₃)₂, t-butyl,cyclopropyl, —C(OH)(CH₃)₂, —OCF₃, —OCHF₂, —O-cyclopropyl,-1-methyl-cyclopropyl, or pyrazolyl.

Provided is also a compound having formula II-b or a pharmaceuticallyacceptable salt or hydrate thereof, wherein R¹, R⁴, and ring A are asdefined in formula I or any one of the above embodiments; R⁷ is H or Cl;and and R¹⁰ is CR¹¹ or N wherein R¹¹ is —F, —SO₂NH₂, —SO₂CH₃, —CN,methoxy, —OCH₂OH, —CH₂OH, —SO₂N(CH₃)₂, —SO₂NHCH₃, —NHSO₂CH₃, —CH₂CH₂OH,—N(CH₃)₂, t-butyl, cyclopropyl, —C(OH)(CH₃)₂, —OCF₃, —OCHF₂,—O-cyclopropyl, -1-methyl-cyclopropyl, or pyrazolyl.

Provided is also a compound having formula II-b-1 or a pharmaceuticallyacceptable salt or hydrate thereof, wherein R¹, R⁴, and ring A are asdefined in formula I or any one of the above embodiments and R^(7′) is Hor Cl.

In another embodiment of formula II, II-a, II-a-1, II-b, or II-b-1,

-   -   R¹ is:

-   -   R⁴ is:

wherein R¹⁰⁰ is H, methyl, Cl, CF₃, CN, OCH₃, or N(R⁶)₂ and R¹⁰¹ is H, For methyl;

-   -   ring A is:

Further embodiments provided herein include combinations of one or moreof the particular embodiments set forth above.

In another embodiment, exemplary compounds of formula I are depictedbelow in Table 1.

Cpd No. Structure 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100

101

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

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Included herein are also methods for making compounds of Formula I or acompound of any one of the embodiments described herein comprisingreacting R¹NC with R²CHO, R³NH₂ and

wherein R^(4′) is H or R⁴ and R¹, R², R³, R⁴ and ring A as defined inFormula I or in any of the embodiments described herein. In one aspectof the preceding methods, R⁴ is alkyl.

Also included herein are methods for making compounds of Formula I or acompound of any one of the embodiments described herein comprising (1)reacting R¹NC with R²CHO, R³NH₂ and

to give

and (2) reacting

with R⁴-halide to give

wherein R⁴ is optionally substituted aryl or optionally substitutedheteroaryl; and R¹, R², R³, R⁴ and ring A as defined in Formula I or inany of the embodiments described herein. In one aspect of the precedingmethods, R⁴ is aryl or heteroaryl, each independently substituted withone to three R⁷ groups. In another aspect of the preceding method, R¹,R², R³, R⁴, R⁵, R⁶, R⁷ and ring A are as defined in any of theembodiments herein.

The compounds of this invention may contain one or more asymmetriccenters and thus occur as racemates, racemic mixtures, scalemicmixtures, and diastereomeric mixtures, as well as single enantiomers orindividual stereoisomers that are substantially free from anotherpossible enantiomer or stereoisomer. The term “substantially free ofother stereoisomers” as used herein means a preparation enriched in acompound having a selected stereochemistry at one or more selectedstereocenters by at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%,96%, 97%, 98%, or 99%. The term “enriched” means that at least thedesignated percentage of a preparation is the compound having a selectedstereochemistry at one or more selected stereocenters. Methods ofobtaining or synthesizing an individual enantiomer or stereoisomer for agiven compound are known in the art and may be applied as practicable tofinal compounds or to starting material or intermediates.

In one embodiment, the compound is enriched in a specific stereoisomerby at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%,or 99%.

The compounds of formula I, II, II-a, II-a-1, II-b or II-b-1 may alsocomprise one or more isotopic substitutions. For example, H may be inany isotopic form, including ¹H, ²H (D or deuterium), and ³H (T ortritium); C may be in any isotopic form, including ¹²C, ¹³C, and ¹⁴C; Omay be in any isotopic form, including ¹⁶O and ¹⁸O; and the like. Forexample, the compound is enriched in a specific isotopic form of H, Cand/or O by at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%,97%, 98%, or 99%.

Unless otherwise indicated when a disclosed compound is named ordepicted by a structure without specifying the stereochemistry and hasone or more chiral centers, it is understood to represent all possiblestereoisomers of the compound.

The compounds of this invention may also be represented in multipletautomeric forms, in such instances, the invention expressly includesall tautomeric forms of the compounds described herein, even though onlya single tautomeric form may be represented (e.g., alkylation of a ringsystem may result in alkylation at multiple sites, the inventionexpressly includes all such reaction products). All such isomeric formsof such compounds are expressly included in the present invention.

Compounds described herein may be prepared following procedures detailedin the examples and other analogous methods known to one skilled in theart. Compounds produced by any of the schemes set forth below may befurther modified (e.g., through the addition of substituents to rings,etc.) to produce additional compounds. The specific approaches andcompounds shown herein are not intended to be limiting. The suitabilityof a chemical group in a compound structure for use in the synthesis ofanother compound is within the knowledge of one of ordinary skill in theart. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing theapplicable compounds are known in the art and include, for example,those described in Larock R, Comprehensive Organic Transformations, VCHPublishers (1989); Greene, T W et al., Protective Groups in OrganicSynthesis, 3^(rd) Ed., John Wiley and Sons (1999); Fieser, L et al.,Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons(1994); and Paquette, L, ed., Encyclopedia of Reagents for OrganicSynthesis, John Wiley and Sons (1995) and subsequent editions thereof.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds.

It may be convenient or desirable to prepare, purify, and/or handle acorresponding salt of the active compound, for example, apharmaceutically acceptable salt. Examples of pharmaceuticallyacceptable salts are discussed in Berge et al., 1977, “PharmaceuticallyAcceptable Salts.” J. Pharm. Sci. Vol. 66, pp. 1-19.

For example, if the compound is anionic, or has a functional group whichmay be anionic (e.g., —COOH may be —COO⁻), then a salt may be formedwith a suitable cation. Examples of suitable inorganic cations include,but are not limited to, alkali metal ions such as Na⁺ and K⁺, alkalineearth cations such as Ca²⁺ and Mg²⁺, and other cations such as Al³⁺.Examples of suitable organic cations include, but are not limited to,ammonium ion (i.e., NH₄ ⁺) and substituted ammonium ions (e.g., NH₃R⁺,NH₂R²⁺, NHR³⁺, NR⁴⁺). Examples of some suitable substituted ammoniumions are those derived from: ethylamine, diethylamine,dicyclohexylamine, triethylamine, butylamine, ethylenediamine,ethanolamine, diethanolamine, piperazine, benzylamine,phenylbenzylamine, choline, meglumine, and tromethamine, as well asamino acids, such as lysine and arginine. An example of a commonquaternary ammonium ion is N(CH₃)₄ ⁺.

If the compound is cationic, or has a functional group that may becationic (e.g., —NH₂ may be —NH₃ ⁺), then a salt may be formed with asuitable anion. Examples of suitable inorganic anions include, but arenot limited to, those derived from the following inorganic acids:hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric,nitrous, phosphoric, and phosphorous.

Examples of suitable organic anions include, but are not limited to,those derived from the following organic acids: 2-acetyoxybenzoic,acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric,edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucoheptonic,gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalenecarboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic,methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic,phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic,succinic, sulfanilic, tartaric, toluenesulfonic, and valeric. Examplesof suitable polymeric organic anions include, but are not limited to,those derived from the following polymeric acids: tannic acid,carboxymethyl cellulose.

Unless otherwise specified, a reference to a particular compound alsoincludes salt forms thereof.

Compositions and Routes of Administration

The compounds utilized in the methods described herein may be formulatedtogether with a pharmaceutically acceptable carrier or adjuvant intopharmaceutically acceptable compositions prior to be administered to asubject. In another embodiment, such pharmaceutically acceptablecompositions further comprise additional therapeutic agents in amountseffective for achieving a modulation of disease or disease symptoms,including those described herein.

The term “pharmaceutically acceptable carrier or adjuvant” refers to acarrier or adjuvant that may be administered to a subject, together witha compound of this invention, and which does not destroy thepharmacological activity thereof and is nontoxic when administered indoses sufficient to deliver a therapeutic amount of the compound.

Pharmaceutically acceptable carriers, adjuvants and vehicles that may beused in the pharmaceutical compositions of this invention include, butare not limited to, ion exchangers, alumina, aluminum stearate,lecithin, self-emulsifying drug delivery systems (SEDDS) such asd-α-tocopherol polyethyleneglycol 1000 succinate, surfactants used inpharmaceutical dosage forms such as Tweens or other similar polymericdelivery matrices, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, potassium sorbate,partial glyceride mixtures of saturated vegetable fatty acids, water,salts or electrolytes, such as protamine sulfate, disodium hydrogenphosphate, potassium hydrogen phosphate, sodium chloride, zinc salts,colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat. Cyclodextrins such as α-, β-, and γ-cyclodextrin, orchemically modified derivatives such as hydroxyalkylcyclodextrins,including 2- and 3-hydroxypropyl-β-cyclodextrins, or other solubilizedderivatives may also be advantageously used to enhance delivery ofcompounds of the formulae described herein.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are mannitol, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose, any bland fixed oil may be employed includingsynthetic mono- or diglycerides. Fatty acids, such as oleic acid and itsglyceride derivatives are useful in the preparation of injectables, asare natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, or carboxymethyl cellulose or similar dispersing agentswhich are commonly used in the formulation of pharmaceuticallyacceptable dosage forms such as emulsions and or suspensions. Othercommonly used surfactants such as Tweens or Spans and/or other similaremulsifying agents or bioavailability enhancers which are commonly usedin the manufacture of pharmaceutically acceptable solid, liquid, orother dosage forms may also be used for the purposes of formulation.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, emulsions and aqueous suspensions,dispersions and solutions. In the case of tablets for oral use, carrierswhich are commonly used include lactose and corn starch. Lubricatingagents, such as magnesium stearate, are also typically added. For oraladministration in a capsule form, useful diluents include lactose anddried corn starch. When aqueous suspensions and/or emulsions areadministered orally, the active ingredient may be suspended or dissolvedin an oily phase is combined with emulsifying and/or suspending agents.If desired, certain sweetening and/or flavoring and/or coloring agentsmay be added.

The pharmaceutical compositions of this invention may also beadministered in the form of suppositories for rectal administration.These compositions can be prepared by mixing a compound of thisinvention with a suitable non-irritating excipient which is solid atroom temperature but liquid at the rectal temperature and therefore willmelt in the rectum to release the active components. Such materialsinclude, but are not limited to, cocoa butter, beeswax and polyethyleneglycols.

Topical administration of the pharmaceutical compositions of thisinvention is useful when the desired treatment involves areas or organsreadily accessible by topical application. For application topically tothe skin, the pharmaceutical composition should be formulated with asuitable ointment containing the active components suspended ordissolved in a carrier. Carriers for topical administration of thecompounds of this invention include, but are not limited to, mineraloil, liquid petroleum, white petroleum, propylene glycol,polyoxyethylene polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical composition can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier with suitable emulsifying agents. Suitablecarriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water. The pharmaceuticalcompositions of this invention may also be topically applied to thelower intestinal tract by rectal suppository formulation or in asuitable enema formulation. Topically-transdermal patches are alsoincluded in this invention.

The pharmaceutical compositions of this invention may be administered bynasal aerosol or inhalation. Such compositions are prepared according totechniques well-known in the art of pharmaceutical formulation and maybe prepared as solutions in saline, employing benzyl alcohol or othersuitable preservatives, absorption promoters to enhance bioavailability,fluorocarbons, and/or other solubilizing or dispersing agents known inthe art. When the compositions of this invention comprise a combinationof a compound of the formulae described herein and one or moreadditional therapeutic or prophylactic agents, both the compound and theadditional agent should be present at dosage levels of between about 1to 100%, and more preferably between about 5 to 95% of the dosagenormally administered in a monotherapy regimen. The additional agentsmay be administered separately, as part of a multiple dose regimen, fromthe compounds of this invention. Alternatively, those agents may be partof a single dosage form, mixed together with the compounds of thisinvention in a single composition.

The compounds described herein can, for example, be administered byinjection, intravenously, intraarterially, subdermally,intraperitoneally, intramuscularly, or subcutaneously; or orally,buccally, nasally, transmucosally, topically, in an ophthalmicpreparation, or by inhalation, with a dosage ranging from about 0.5 toabout 100 mg/kg of body weight, alternatively dosages between 1 mg and1000 mg/dose, every 4 to 120 hours, or according to the requirements ofthe particular drug. The methods herein contemplate administration of aneffective amount of compound or compound composition to achieve thedesired or stated effect. Typically, the pharmaceutical compositions ofthis invention will be administered from about 1 to about 6 times perday or alternatively, as a continuous infusion. Such administration canbe used as a chronic or acute therapy. The amount of active ingredientthat may be combined with the carrier materials to produce a singledosage form will vary depending upon the host treated and the particularmode of administration. A typical preparation will contain from about 5%to about 95% active compound (w/w). Alternatively, such preparationscontain from about 20% to about 80% active compound.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular subject will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the subject'sdisposition to the disease, condition or symptoms, and the judgment ofthe treating physician.

Upon improvement of a subject's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level. Subjects may, however,require intermittent treatment on a long-term basis upon any recurrenceof disease symptoms.

The pharmaceutical compositions described above comprising a compound offormula I, II, II-a, II-a-1, II-b, or II-b-1 or a compound described inany one of the embodiments herein, may further comprise anothertherapeutic agent useful for treating cancer.

Methods of Use

Provided is a method for inhibiting a mutant IDH1 or IDH2 activitycomprising contacting a subject in need thereof with a compound(including its tautomers and/or isotopologues) of structural formula I,II, II-a, II-a-1, II-b, or II-b-1 or a compound described in any one ofthe embodiments herein, or a pharmaceutically acceptable salt thereof.In one embodiment, the cancer to be treated is characterized by a mutantallele of IDH1 or IDH2 wherein the IDH1 or IDH2 mutation results in anew ability of the enzyme to catalyze the NAPH-dependent reduction ofα-ketoglutarate to R(−)-2-hydroxyglutarate in a subject. In one aspectof this embodiment, the mutant IDH1 has an R132X mutation. In one aspectof this embodiment, the R132X mutation is selected from R132H, R132C,R132L, R132V, R132S and R132G. In another aspect, the R132X mutation isR132H or R132C. In yet another aspect, the R132X mutation is R132H.

Also provided are methods of treating a cancer characterized by thepresence of a mutant allele of IDH1 comprising the step of administeringto subject in need thereof (a) a compound of formula I, II, II-a,II-a-1, II-b, or II-b-1, or a compound described in any one of theembodiments herein, or a pharmaceutically acceptable salt thereof, or(b) a pharmaceutical composition comprising (a) and a pharmaceuticallyacceptable carrier.

In one embodiment, the cancer to be treated is characterized by a mutantallele of IDH1 wherein the IDH1 mutation results in a new ability of theenzyme to catalyze the NAPH-dependent reduction of α-ketoglutarate toR(−)-2-hydroxyglutarate in a patient. In one aspect of this embodiment,the IDH1 mutation is an R132X mutation. In another aspect of thisembodiment, the R132X mutation is selected from R132H, R132C, R132L,R132V, R132S and R132G. In another aspect, the R132X mutation is R132Hor R132C. A cancer can be analyzed by sequencing cell samples todetermine the presence and specific nature of (e.g., the changed aminoacid present at) a mutation at amino acid 132 of IDH1.

Without being bound by theory, applicants believe that mutant alleles ofIDH1 wherein the IDH1 mutation results in a new ability of the enzyme tocatalyze the NAPH-dependent reduction of α-ketoglutarate toR(−)-2-hydroxyglutarate, and in particular R132H mutations of IDH1,characterize a subset of all types of cancers, without regard to theircellular nature or location in the body. Thus, the compounds and methodsof this invention are useful to treat any type of cancer that ischaracterized by the presence of a mutant allele of IDH1 imparting suchacitivity and in particular an IDH1 R132H or R132C mutation.

In one aspect of this embodiment, the efficacy of cancer treatment ismonitored by measuring the levels of 2HG in the subject. Typicallylevels of 2HG are measured prior to treatment, wherein an elevated levelis indicated for the use of the compound of formula I, II, II-a, II-a-1,II-b, or II-b-1 or a compound described in any one of the embodimentsdescribed herein to treat the cancer. Once the elevated levels areestablished, the level of 2HG is determined during the course of and/orfollowing termination of treatment to establish efficacy. In certainembodiments, the level of 2HG is only determined during the course ofand/or following termination of treatment. A reduction of 2HG levelsduring the course of treatment and following treatment is indicative ofefficacy. Similarly, a determination that 2HG levels are not elevatedduring the course of or following treatment is also indicative ofefficacy. Typically, the these 2HG measurements will be utilizedtogether with other well-known determinations of efficacy of cancertreatment, such as reduction in number and size of tumors and/or othercancer-associated lesions, improvement in the general health of thesubject, and alterations in other biomarkers that are associated withcancer treatment efficacy.

2HG can be detected in a sample by LC/MS. The sample is mixed 80:20 withmethanol, and centrifuged at 3,000 rpm for 20 minutes at 4 degreesCelsius. The resulting supernatant can be collected and stored at −80degrees Celsius prior to LC-MS/MS to assess 2-hydroxyglutarate levels. Avariety of different liquid chromatography (LC) separation methods canbe used. Each method can be coupled by negative electrospray ionization(ESI, −3.0 kV) to triple-quadrupole mass spectrometers operating inmultiple reaction monitoring (MRM) mode, with MS parameters optimized oninfused metabolite standard solutions. Metabolites can be separated byreversed phase chromatography using 10 mM tributyl-amine as an ionpairing agent in the aqueous mobile phase, according to a variant of apreviously reported method (Luo et al. J Chromatogr A 1147, 153-64,2007). One method allows resolution of TCA metabolites: t=0, 50% B; t=5,95% B; t=7, 95% B; t=8, 0% B, where B refers to an organic mobile phaseof 100% methanol. Another method is specific for 2-hydroxyglutarate,running a fast linear gradient from 50%-95% B (buffers as defined above)over 5 minutes. A Synergi Hydro-RP, 100 mm×2 mm, 2.1 μm particle size(Phenomonex) can be used as the column, as described above. Metabolitescan be quantified by comparison of peak areas with pure metabolitestandards at known concentration. Metabolite flux studies from¹³C-glutamine can be performed as described, e.g., in Munger et al. NatBiotechnol 26, 1179-86, 2008.

In one embodiment 2HG is directly evaluated.

In another embodiment a derivative of 2HG formed in process ofperforming the analytic method is evaluated. By way of example such aderivative can be a derivative formed in MS analysis. Derivatives caninclude a salt adduct, e.g., a Na adduct, a hydration variant, or ahydration variant which is also a salt adduct, e.g., a Na adduct, e.g.,as formed in MS analysis.

In another embodiment a metabolic derivative of 2HG is evaluated.Examples include species that build up or are elevated, or reduced, as aresult of the presence of 2HG, such as glutarate or glutamate that willbe correlated to 2HG, e.g., R-2HG.

Exemplary 2HG derivatives include dehydrated derivatives such as thecompounds provided below or a salt adduct thereof:

In one embodiment the cancer is a tumor wherein at least 30, 40, 50, 60,70, 80 or 90% of the tumor cells carry an IDH1 mutation, and inparticular an IDH1 R132H or R132C mutation, at the time of diagnosis ortreatment.

IDH1 R132X mutations are known to occur in certain types of cancers asindicated in Table 2, below.

TABLE 2 IDH mutations associated with certain cancers IDH1 R132X CancerType Mutation Tumor Type brain tumors R132H primary tumor R132C primarytumor R132S primary tumor R132G primary tumor R132L primary tumor R132Vprimary tumor fibrosarcoma R132C HT1080 fibrosarcoma cell line AcuteMyeloid Leukemia R132H primary tumor (AML) R132G primary tumor R132Cprimary tumor Prostate cancer R132H primary tumor R132C primary tumorAcute lymphoblastic leukemia R132C primary tumor (ALL) paragangliomasR132C primary tumor

IDH1 R132H mutations have been identified in glioblastoma, acutemyelogenous leukemia, sarcoma, melanoma, non-small cell lung cancer,cholangiocarcinomas, chondrosarcoma, myelodysplastic syndromes (MDS),myeloproliferative neoplasm (MPN), colon cancer, and angio-immunoblasticnon-Hodgkin's lymphoma (NHL). Accordingly, in one embodiment, themethods described herein are used to treat glioma (glioblastoma), acutemyelogenous leukemia, sarcoma, melanoma, non-small cell lung cancer(NSCLC) or cholangiocarcinomas, chondrosarcoma, myelodysplasticsyndromes (MDS), myeloproliferative neoplasm (MPN), colon cancer, orangio-immunoblastic non-Hodgkin's lymphoma (NHL) in a patient.

Accordingly in one embodiment, the cancer is a cancer selected from anyone of the cancer types listed in Table 2, and the IDH R132X mutation isone or more of the IDH1 R132X mutations listed in Table 2 for thatparticular cancer type.

Treatment methods described herein can additionally comprise variousevaluation steps prior to and/or following treatment with a compound offormula I, II, II-a, II-a-1, II-b, or II-b-1 or a compound described inany one of the embodiments described herein.

In one embodiment, prior to and/or after treatment with a compound ofStructural formula I, II, II-a, II-a-1, II-b, or II-b-1 or a compounddescribed in any one of the embodiments described herein, the methodfurther comprises the step of evaluating the growth, size, weight,invasiveness, stage and/or other phenotype of the cancer.

In one embodiment, prior to and/or after treatment with a compound offormula I, II, II-a, II-a-1, II-b, or II-b-1 or a compound described inany one of the embodiments described herein, the method furthercomprises the step of evaluating the IDH1 genotype of the cancer. Thismay be achieved by ordinary methods in the art, such as DNA sequencing,immuno analysis, and/or evaluation of the presence, distribution orlevel of 2HG.

In one embodiment, prior to and/or after treatment with a compound offormula I, II, II-a, II-a-1, II-b, or II-b-1 or a compound described inany one of the embodiments described herein, the method furthercomprises the step of determining the 2HG level in the subject. This maybe achieved by spectroscopic analysis, e.g., magnetic resonance-basedanalysis, e.g., MRI and/or MRS measurement, sample analysis of bodilyfluid, such as serum or spinal cord fluid analysis, or by analysis ofsurgical material, e.g., by mass-spectroscopy.

Combination Therapies

In some embodiments, the methods described herein comprise theadditional step of co-administering to a subject in need thereof asecond therapy e.g., an additional cancer therapeutic agent or anadditional cancer treatment. Exemplary additional cancer therapeuticagents include for example, chemotherapy, targeted therapy, antibodytherapies, immunotherapy, and hormonal therapy. Additional cancertreatments include, for example: surgery, and radiation therapy.Examples of each of these treatments are provided below.

The term “co-administering” as used herein with respect to an additionalcancer therapeutic agents means that the additional cancer therapeuticagent may be administered together with a compound of this invention aspart of a single dosage form (such as a composition of this inventioncomprising a compound of the invention and an second therapeutic agentas described above) or as separate, multiple dosage forms.Alternatively, the additional cancer therapeutic agent may beadministered prior to, consecutively with, or following theadministration of a compound of this invention. In such combinationtherapy treatment, both the compounds of this invention and the secondtherapeutic agent(s) are administered by conventional methods. Theadministration of a composition of this invention, comprising both acompound of the invention and a second therapeutic agent, to a subjectdoes not preclude the separate administration of that same therapeuticagent, any other second therapeutic agent or any compound of thisinvention to said subject at another time during a course of treatment.The term “co-administering” as used herein with respect to an additionalcancer treatment means that the additional cancer treatment may occurprior to, consecutively with, concurrently with or following theadministration of a compound of this invention.

In some embodiments, the additional cancer therapeutic agent is achemotherapy agent. Examples of chemotherapeutic agents used in cancertherapy include, for example, antimetabolites (e.g., folic acid, purine,and pyrimidine derivatives), alkylating agents (e.g., nitrogen mustards,nitrosoureas, platinum, alkyl sulfonates, hydrazines, triazenes,aziridines, spindle poison, cytotoxic agents, topoisomerase inhibitorsand others) and hypomethylating agents (e.g., decitabine(5-aza-deoxycytidine), zebularine, isothiocyanates, azacitidine(5-azacytidine, 5-flouro-2′-deoxycytidine, 5,6-dihydro-5-azacytidine andothers). Exemplary agents include Aclarubicin, Actinomycin,Alitretinoin, Altretamine, Aminopterin, Aminolevulinic acid, Amrubicin,Amsacrine, Anagrelide, Arsenic trioxide, Asparaginase, Atrasentan,Belotecan, Bexarotene, bendamustine, Bleomycin, Bortezomib, Busulfan,Camptothecin, Capecitabine, Carboplatin, Carboquone, Carmofur,Carmustine, Celecoxib, Chlorambucil, Chlormethine, Cisplatin,Cladribine, Clofarabine, Crisantaspase, Cyclophosphamide, Cytarabine,Dacarbazine, Dactinomycin, Daunorubicin, Decitabine, Demecolcine,Docetaxel, Doxorubicin, Efaproxiral, Elesclomol, Elsamitrucin,Enocitabine, Epirubicin, Estramustine, Etoglucid, Etoposide,Floxuridine, Fludarabine, Fluorouracil (5FU), Fotemustine, Gemcitabine,Gliadel implants, Hydroxycarbamide, Hydroxyurea, Idarubicin, Ifosfamide,Irinotecan, Irofulven, Ixabepilone, Larotaxel, Leucovorin, Liposomaldoxorubicin, Liposomal daunorubicin, Lonidamine, Lomustine, Lucanthone,Mannosulfan, Masoprocol, Melphalan, Mercaptopurine, Mesna, Methotrexate,Methyl aminolevulinate, Mitobronitol, Mitoguazone, Mitotane, Mitomycin,Mitoxantrone, Nedaplatin, Nimustine, Oblimersen, Omacetaxine, Ortataxel,Oxaliplatin, Paclitaxel, Pegaspargase, Pemetrexed, Pentostatin,Pirarubicin, Pixantrone, Plicamycin, Porfimer sodium, Prednimustine,Procarbazine, Raltitrexed, Ranimustine, Rubitecan, Sapacitabine,Semustine, Sitimagene ceradenovec, Strataplatin, Streptozocin,Talaporfin, Tegafur-uracil, Temoporfin, Temozolomide, Teniposide,Tesetaxel, Testolactone, Tetranitrate, Thiotepa, Tiazofurine,Tioguanine, Tipifarnib, Topotecan, Trabectedin, Triaziquone,Triethylenemelamine, Triplatin, Tretinoin, Treosulfan, Trofosfamide,Uramustine, Valrubicin, Verteporfin, Vinblastine, Vincristine,Vindesine, Vinflunine, Vinorelbine, Vorinostat, Zorubicin, and othercytostatic or cytotoxic agents described herein.

Because some drugs work better together than alone, two or more drugsare often given at the same time. Often, two or more chemotherapy agentsare used as combination chemotherapy.

In some embodiments, the additional cancer therapeutic agent is adifferentiation agent. Such differentiation agent includes retinoids(such as all-trans-retinoic acid (ATRA), 9-cis retinoic acid,13-cis-retinoic acid (13-cRA) and 4-hydroxy-phenretinamide (4-HPR));arsenic trioxide; histone deacetylase inhibitors HDACs (such asazacytidine (Vidaza) and butyrates (e.g., sodium phenylbutyrate));hybrid polar compounds (such as hexamethylene bisacetamide ((HMBA));vitamin D; and cytokines (such as colony-stimulating factors includingG-CSF and GM-CSF, and interferons).

In some embodiments the additional cancer therapeutic agent is atargeted therapy agent. Targeted therapy constitutes the use of agentsspecific for the deregulated proteins of cancer cells. Small moleculetargeted therapy drugs are generally inhibitors of enzymatic domains onmutated, overexpressed, or otherwise critical proteins within the cancercell. Prominent examples are the tyrosine kinase inhibitors such asAxitinib, Bosutinib, Cediranib, dasatinib, erlotinib, imatinib,gefitinib, lapatinib, Lestaurtinib, Nilotinib, Semaxanib, Sorafenib,Sunitinib, and Vandetanib, and also cyclin-dependent kinase inhibitorssuch as Alvocidib and Seliciclib. Monoclonal antibody therapy is anotherstrategy in which the therapeutic agent is an antibody whichspecifically binds to a protein on the surface of the cancer cells.Examples include the anti-HER2/neu antibody trastuzumab (HERCEPTIN®)typically used in breast cancer, and the anti-CD20 antibody rituximaband Tositumomab typically used in a variety of B-cell malignancies.Other exemplary antibodies include Cetuximab, Panitumumab, Trastuzumab,Alemtuzumab, Bevacizumab, Edrecolomab, and Gemtuzumab. Exemplary fusionproteins include Aflibercept and Denileukin diftitox. In someembodiments, the targeted therapy can be used in combination with acompound described herein, e.g., a biguanide such as metformin orphenformin, preferably phenformin.

Targeted therapy can also involve small peptides as “homing devices”which can bind to cell surface receptors or affected extracellularmatrix surrounding the tumor. Radionuclides which are attached to thesepeptides (e.g., RGDs) eventually kill the cancer cell if the nuclidedecays in the vicinity of the cell. An example of such therapy includesBEXXAR®.

In some embodiments, the additional cancer therapeutic agent is animmunotherapy agent. Cancer immunotherapy refers to a diverse set oftherapeutic strategies designed to induce the subject's own immunesystem to fight the tumor. Contemporary methods for generating an immuneresponse against tumors include intravesicular BCG immunotherapy forsuperficial bladder cancer, and use of interferons and other cytokinesto induce an immune response in renal cell carcinoma and melanomasubjects.

Allogeneic hematopoietic stem cell transplantation can be considered aform of immunotherapy, since the donor's immune cells will often attackthe tumor in a graft-versus-tumor effect. In some embodiments, theimmunotherapy agents can be used in combination with a compound orcomposition described herein.

In some embodiments, the additional cancer therapeutic agent is ahormonal therapy agent. The growth of some cancers can be inhibited byproviding or blocking certain hormones. Common examples ofhormone-sensitive tumors include certain types of breast and prostatecancers. Removing or blocking estrogen or testosterone is often animportant additional treatment. In certain cancers, administration ofhormone agonists, such as progestogens may be therapeuticallybeneficial. In some embodiments, the hormonal therapy agents can be usedin combination with a compound or a composition described herein.

Other possible additional therapeutic modalities include imatinib, genetherapy, peptide and dendritic cell vaccines, synthetic chlorotoxins,and radiolabeled drugs and antibodies.

EXAMPLES

The chemical name of each compound described below is generated byChemBioOffice software.

DCM=dichloromethane TEA=triethylamine

DPPA=diphenyiphosphoryl azide TFA=trifluoroacetic acid

DIPEA=N,N-Diisopropylethylamine TFAA=trifluoroacetic anhydride

General Procedures for the Preparation of1,1-difluoro-3-isocyanocyclobutane

Step A: Tert-butyl 3-oxocyclobutykarbamate

To a solution of 3-oxocyclobutanecarboxylic acid (10 g, 88 mmol) in dryDCM (60 mL) at 0° C., SOCl₂ (20 mL) was added dropwise. The mixture washeated to reflux for 1.5 h and then evaporated in vacuo. The resultingmixture was co-evaporated twice with toluene (2×8 mL) and the residuewas dissolved in acetone (30 mL), followed by adding dropwise to asolution of NaN₃ (12 g, 185.0 mmol) in H₂O (35 mL) at 0° C. Afteraddition, the mixture was stirred for another 1 h and then quenched withice (110 g). The resulting mixture was extracted with Et₂O (2×100 mL).Combined organic layers were washed with brine, dried over anhydrousMg₂SO₄ and concentrated to about 15 mL solution. Toluene (2×30 mL) wasadded into the residue and the mixture was co-evaporated twice to removeEt₂O (about 30 mL solution left each time to avoid explosion). Theresulting toluene solution was heated to 90° C. until the evolution ofN₂ ceased. Next, 40 mL of t-BuOH was added into the reaction mixture andthe resulting mixture was stirred overnight at 90° C. The mixture wascooled and concentrated. The residue was purified by columnchromatography using petroleum ether/EtOAc (V:V, 7:1 to 5:1) as eluentto afford the desired product as a white solid. MS: 186.1 (M+1)⁺.

Step B: Tert-butyl 3,3-difluorocyclobutylcarbamate

To a solution of tert-butyl-3-oxocyclo-butylcarbamate (2.56 g, 111.07mmol) in dry DCM (190 mL), DAST (diethylaminosulfur trifluoride) (41.0mL, 222.14 mmol) was added dropwise at 0° C. under the atmosphere of N₂.The mixture was then allowed to warm up to r.t and stirred overnight.The resulting mixture was slowly added into a pre-cooled saturated aq.NaHCO₃ solution and extracted with DCM (3×200 mL). Combined organiclayers were washed with brine, dried over anhydrous MgSO₄, andconcentrated in vacuo. The residue was purified by column chromatographyusing petroleum ether/EtOAc (V:V, 15:1) as eluent to afford the desiredproduct. ¹H NMR (400 MHz, DMSO-d₆): δ 4.79 (s, 1H), 4.07 (s, 1H), 2.98(s, 2H), 2.58-2.29 (m, 2H), 1.46 (s, 9H). MS: 208.1 (M+1)⁺.

Step C: N-(3,3-difluorocyclobutyl)formamide

To a solution of MeOH (170 mL) and CH₃COCl (65 mL), tert-butyl3,3-difluoro-cyclobutylcarbamate (12.1 g, 58.42 mmol) was added in oneportion dropwise at 0° C. The reaction mixture was stirred at 0° C. for20 min, and then allowed to warm up to r.t and stirred for another 1.5h. The reaction mixture was concentrated and dissolved in H₂O (200 mL).The resulting mixture was extracted by Et₂O (150 mL) and the aqueouslayer was adjusted to pH=11 with solid Na₂CO₃ and extracted by DCM(2×150 mL). The combined organic layers were dried over anhydrous MgSO₄,filtered and concentrated in vacuo using a cold-water bath (<20° C.).The residue was dissolved in HCOOEt (90 mL), and transferred into asealed pressure tube. This reaction mixture was heated to 80° C. andstirred overnight. The solvent was removed, and the residue was purifiedby column chromatography using petroleum ether/EtOAc (V:V, 1:1 to 1:3)as eluent to afford the desired product. MS: 136.1 (M+1)⁺.

Step D: 1,1-Difluoro-3-isocyanocyclobutane

To a solution of N-(3,3-difluorocyclobutyl)-formamide (2.0 g, 14.81mmol) and PPh₃ (4.27 g, 16.29 mmol) in DCM (35 mL) were added CCl₄(1.43mL, 14.81 mmol) and TEA (2.06 mL, 14.81 mmol). The reaction mixture wasstirred at 45° C. overnight under a N₂ atmosphere. The resulting mixturewas evaporated in vacuo at 0° C. The residue was suspended in Et₂O (25mL) at 0° C. for 30 min and then filtered. The filtrate was evaporatedto about 5 mL at 0° C. under reduced pressure. The residue was purifiedby column chromatography using Et₂O as eluent to afford the desiredproduct which was used directly in the next step.

Step A: Benzyl 3-oxocyclobutanecarboxylate

A mixture of 3-oxocyclobutanecarboxylic acid (5 g, 44 mmol), potassiumcarbonate (12 g, 88 mmol) and benzyl bromide (11.2 g, 66 mmol) inacetone (50 mL) was refluxed for 16 h. The solvent was then removedunder reduced pressure and the residue was partitioned between ethylacetate and water. Combined organic layers were dried over anhydrousMgSO₄, filtered and concentrated. The residue was purified with silicagel chromatography eluting with a gradient of 100% hexane to 96%hexane/EtOAc to give the desired compound. ¹H NMR (400 MHz, CDCl₃): δ7.45-7.27 (m, 5H), 5.19 (s, 2H), 3.55-3.36 (m, 2H), 3.33-3.11 (m, 3H).

Step B: Benzyl 3,3-difluorocyclobutanecarboxylate

To a solution of benzyl 3-oxocyclobutanecarboxylate (1.23 g, 6.03 mmol)in DCM (35 mL) was added DAST (0.8 mL, 6.03 mmol) dropwise undernitrogen. The mixture was stirred at room temperature for 16 h and thendiluted with DCM. After successive washes with saturated sodiumbicarbonate, 1N aq. hydrochloride acid, and brine, the organic layer wasdried over anhydrous sodium sulfate, filtered and concentrated. Thecrude product was purified by silica gel chromatography with 93%hexane/EtOAc as eluent to give the desired compound as an oil. ¹H NMR(400 MHz, CDCl₃): δ 7.47-7.27 (m, 5H), 5.16 (s, 2H), 3.09-2.95 (m, 1H),2.90-2.60 (m, 4H).

Step C: 3,3-Difluorocyclobutanecarboxylic acid

Benzyl 3,3-difluorocyclobutanecarboxylate (0.84 g, 3.72 mol) wasdissolved in ethanol (40 mL), and approximately 0.02 g palladium onactivated carbon was added. The mixture was stirred at room temperaturefor 12 h under the atmosphere of H₂ and then filtered through a pad ofCelite. The filtrates were concentrated and dried in vacuo to give thedesired compound. ¹H NMR (400 MHz, CDCl₃): δ 3.16-2.55 (m, 5H).

Step D: Tert-butyl 3,3-difluorocyclobutylcarbamate

Benzyl 3,3-difluorocyclobutanecarboxylic acid (3.7 g, 27.3 mmol), DPPA(7.87 g, 27 mmol) and TEA (2.87 g, 28.4 mmol) were dissolved in t-BuOH(25 mL). The mixture was refluxed for 5 h and then diluted with ethylacetate (about 200 mL). The organic phase was washed twice with 5%citric acid and saturated sodium hydrogen carbonate respectively, driedover anhydrous Mg₂SO₄ and evaporated under reduced pressure. The residuewas purified by silica gel chromatography with 50% hexane/EtOAc to givethe desired product. MS: 208.1 (M+1)⁺.

Step E: 3,3-Difluorocyclobutanamine hydrochloride

To a cold solution of MeOH (170 mL) and CH₃COCl (65 mL) was addedtert-butyl 3,3-difluorocyclobutylcarbamate (12.1 g, 58.4 mmol) dropwiseat 0° C. After completion of the addition, the mixture was stirred at 0°C. for 20 min and then allowed to warm up to room temperature. Thereaction mixture was stirred for another 1.5 h and then concentrated togive the crude product which was precipitated in ether to give thedesired product as a white solid. MS: 108.1 (M+1)⁺.

Step F: N-(3,3-difluorocyclobutyl)formamide

The mixture of 3,3-difluorocyclobutanamine hydrochloride (6.5 g, 60.7mmol) and TEA (3 eq) in HCOOEt (90 mL) was stirred at 80° C. overnightin a sealed pressure tube. The solvent was removed in vacuo and theresidue was purified by column chromatography with 50% petroleumether/EtOAc to 25% petroleum ether/EtOAc to give the desired product. ¹HNMR (400 MHz, DMSO-d₆): δ 8.54 (s, 1H), 8.01-7.89 (m, 1H), 4.16-3.84 (m,1H), 3.06-2.73 (m, 2H), 2.72-2.33 (m, 2H). MS: 136.1 (M+1)⁺.

Step G: 1,1-Difluoro-3-isocyanocyclobutane

The compound was synthesized as outlined in step D of method A set forthabove.

General Procedures for the Preparation of 1-fluoro-3-isocyanocyclobutane

Step A: Tert-butyl 3-hydroxycyclobutylcarbamate

To a solution of tert-butyl 3-oxocyclobutylcarbamate (2 g, 10.8 mmol, 2eq) in EtOH (20 mL) was added NaBH₄ (204 mg, 1 eq) at 0° C. The mixturewas then allowed to warm to room temperature and stirred for 30 min. Themixture was concentrated in vacuo and the residue was purified by columnchromatography using petroleum ether/EtOAc (V:V, 2:1 to pure EtOAc) aseluent to afford the desired product as a white solid. MS: 188.1 (M+1)⁺.

Step B: Tert-butyl 3-fluorocyclobutykarbamate

To a solution tert-butyl 3-hydroxycyclobutyl carbamate (1 g, 5.35 mmol)in dry DCM (20 mL) at −70° C. was added DAST dropwise (1 g, 0.85 mL,1.17 eq) under the atmosphere of N₂. The mixture was then slowly warmedto room temperature and stirred overnight. The resulting mixture waswashed with diluted aq. NaHCO₃. The organic layer was dried overanhydrous Mg₂SO₄ and concentrated. The residue was purified by flashchromatography using petroleum ether/EtOAc (V:V, 20:1 to 2:1) as eluentto afford a white solid as the desired product. MS: 190.1 (M+1)⁺.

Step C: 3-Fluorocyclobutanamine

The compound was synthesized as outlined in step E of method A set forthabove.

Step D: N-(3-fluorocyclobutyl)formamide

The compound was synthesized as outlined in step F of method A set forthabove. ¹H NMR (400 MHz, CDCl₃): δ 8.10 (s, 1H), 5.94-5.89 (brs, 1H),5.32-5.25 (m, 0.5H), 5.18-5.11 (m, 0.5H), 4.63-4.42 (m, 1H), 2.76-2.62(m, 2H), 2.44-2.31 (m, 2H).

Step E: 1-Fluoro-3-isocyanocyclobutane

The compound was synthesized via the general procedure as the step G inmethod A set forth above.

General Procedures for the Preparation of1,1-difluoro-4-isocyanocyclohexane

Step A: Tert-butyl 4-hydroxycyclohexylcarbamate

To a solution of 4-aminocyclohexanol (23 g, 0.2 mol) and Et₃N (60 g, 0.6mol) in THF (230 mL) was added (Boc)₂O (87 g, 0.4 mol). The resultingsolution was stirred at room temperature overnight. The solvent wasremoved under reduced pressure and the residue was extracted with EtOAc(3×200 mL). The combined organic layers were washed with water (2×200mL) and brine (200 mL), dried over anhydrous Na₂SO₄ and concentrated.The residue was purified by column chromatography on silica gel usingDCM/MeOH (V:V, 20:1) to afford the desired product as a white solid. MS:216.2 (M+1)⁺.

Step B: Tert-butyl 4-oxocyclohexylcarbamate

To a solution of tert-butyl 4-hydroxycyclohexyl-carbamate (10.0 g, 46.5mmol) in DCM (100 mL) was added Dess-Martin periodinane (39.4 g, 92.9mmol) portionwise. The resulting solution was stirred at roomtemperature overnight, quenched with aq. Na₂S₂O₃ solution and extractedwith DCM (3×100 mL). The combined organic layers were washed with water(2×100 mL) and brine (100 mL), dried over anhydrous Na₂SO₄, andconcentrated. The residue was purified by column chromatography onsilica gel using petroleum ether/EtOAc (V:V, 10:1) to afford desiredproduct as a white solid.

Step C: Tert-butyl 4,4-difluorocyclohexylcarbamate

To a solution of tert-butyl 4-oxocyclo-hexylcarbamate (2.13 g, 10 mmol)in dry DCM (25 mL) was added DAST (2.58 g, 16 mmol) dropwise at −5° C.under nitrogen. After addition, the reaction mixture was stirred at r.tovernight. The reaction mixture was poured into ice water slowly andextracted with DCM (3×100 mL). The combined organic layers were washedwith 2 N aq. NaHCO₃ and brine, dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by column chromatographyusing petroleum ether/EtOAc (V:V, 5:1) as eluent to afford a mixture ofthe title compound (˜70%) and the byproduct tert-butyl4-fluorocyclohex-3-enylcarbamate (˜30%) as a light-yellow solid.

To the above mixtures (2.52 g, 10.7 mmol) in DCM (25 mL) was addedm-CPBA (2.20 g, 12.9 mmol) portionwise at 0° C. while keeping theinternal temperature below 5° C. After addition, the reaction mixturewas stirred at room temperature overnight. To the reaction mixture wasadded saturated aq. Na₂S₂O₃ (8.0 mL) at 0° C. The resulting mixture wasstirred at 0° C. for 40 min, and then extracted by DCM (3×5.0 mL). Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄, and evaporated in vacuo. The residue was used directly in thenext step without further purification.

To the above residue in MeOH (15 mL) was added NaBH₄ (0.202 g, 5.35mmol) at 0° C. The reaction mixture was stirred at room temperatureovernight. Water (0.38 g) was added dropwise to quench the reaction at0° C. The resulting mixture was stirred at 0° C. for 30 min, andconcentrated in vacuo. The residue was purified by column chromatographyusing DCM as eluent to afford the pure compound as a white solid. ¹H NMR(400 MHz, CDCl₃): δ 4.46 (s, 1H), 3.59 (s, 1H), 2.25-1.69 (m, 6H),1.61-1.20 (m, 11H). MS: 236.2 (M+1)⁺.

Step D: 4,4-Difluorocyclohexanamine hydrochloride

A mixture of tert-butyl 4,4-difluorocyclo-hexylcarbamate (6.0 g, 25.5mmol) and 6 N HCl/MeOH (60 mL) was stirred at room temperature for 2 h.The reaction mixture was concentrated to give the crude product whichwas directly used in next step without further purification. ¹H NMR (400MHz, CD₃OD): δ 4.89 (s, 2H), 3.32-3.26 (m, 1H), 2.14-2.01 (m, 4H),2.02-1.85 (m, 2H), 1.74-1.65 (m, 2H). MS: 136.1 (M+1)⁺.

Step E: N-(4,4-difluorocyclohexyl)formamide

A mixture of 4,4-difluorocyclohexanamine (crude 3.4 g, 25.2 mmol), TEA(3 eq) and ethyl formate (35 mL) was stirred at 110° C. overnight in asealed tank. The solvent was removed and the residue was purified bycolumn chromatography using DCM/MeOH (V:V, 10:1) as eluent to afford thedesired product. ¹H NMR (400 MHz, CDCl₃): δ 8.14 (s, 1H), 5.98 (s, 1H),3.93 (m, 1H), 2.54-2.19 (m, 1H), 2.15-1.39 (m, 7H). MS: 164.1 (M+1)⁺.

Step F: 1,1-Difluoro-4-isocyanocyclohexane

A mixture of N-(4,4-difluorocyclohexyl)-formamide (2.5 g, 15.3 mmol),PPh₃ (4.4 g, 16.8 mmol), CCl₄ (2.3 g, 15.1 mmol), Et₃N (1.5 g, 14.9mmol) and DCM (50 mL) was heated to 45° C. and stirred overnight. Theresulting mixture was evaporated in vacuo and the residue was suspendedin Et₂O (125 mL) at 0° C. The filtrate was concentrated and the residuewas purified by column chromatography on silica gel eluting with Et₂O toafford the desired product as a yellow oil which was used directly inthe next step.

General Procedures for the Preparation of 2-(3-aminophenoxy)ethanol

Step A: 2-(3-Nitrophenoxy)ethanol

A suspension of 3-nitrophenol (1 g, 7.2 mmol), 2-bromoethanol (1.2 g,9.6 mmol) and K₂CO₃ (2 g, 14.4 mmol) in MeCN (12 mL) was stirred at 90°C. overnight. The precipitate was collected by filtration to give thefirst batch of product. The filtrate was concentrated and the residuewas purified by column chromatography to afford another batch of thedesired product as a yellow solid.

Step B: 2-(3-Aminophenoxy)ethanol

To a solution of 2-(3-nitrophenoxy)ethanol (500 mg, 2.7 mmol) and NH₄Cl(720 mg, 13.5 mmol) in EtOH (10 mL) was added iron powder (900 mg, 16.2mmol) at room temperature. The reaction was then stirred at 90° C. for 2hr and subsequently cooled. The mixture was filtered and the filtratewas concentrated. The resulting residue was purified by columnchromatography to afford the desired product as a yellow solid. MS:154.1 (M+1)⁺.

General Procedures for the Preparation of 3-(1H-pyrazol-4-yl)aniline

Step A: Tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-1-carboxylate

To a solution of4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (500 mg,2.57 mmol) and (Boc)₂O (672 mg, 3.08 mmol) in DMF (1.0 mL) was addedDMAP (63 mg, 0.52 mmol) in one portion. The mixture was stirred at roomtemperature overnight, and then partitioned between EtOAc and saturatedaq. NH₄Cl. The organic layer was separated, washed with brine, driedover anhydrous Na₂SO₄, and concentrated to afford the crude product.

Step B: 4-(3-Nitrophenyl)-1H-pyrazole

To a solution of tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-1-carboxylate(300 mg, 0.82 mmol), 1-bromo-3-nitrobenzene (137 mg, 0.68 mmol) andNa₂CO₃ (216 mg, 2.04 mmol) in DME/H₂O (5 mL/1 mL) under N₂, was addedPd(PPh₃)₂Cl₂ (24 mg, 0.034 mmol). The mixture was stirred at 85° C.overnight, and then quenched with H₂O. The resulting mixture wasextracted with EtOAc (3×25 mL). The organic layer was separated, washedwith brine, dried over anhydrous Na₂SO₄, and concentrated. The resultingresidue was purified by column chromatography to afford the desiredproduct. MS: 190.2 (M+1)⁺.

Step C: 3-(1H-pyrazol-4-yl)aniline

Iron powder (296 mg, 5.30 mmol) was added to a solution of4-(3-nitrophenyl)-1H-pyrazole (200 mg, 1.06 mmol) in AcOH/EtOH (2 mL/3mL). The reaction mixture was stirred at 90° C. for 2 hr and then cooledto room temperature. The reaction mixture was filtered through Celite.The filter cake was washed with H₂O. The filtrate was neutralized with 1N NaOH to pH=8 and extracted with EtOAc (3×30 mL). The combined organiclayers were washed with brine, dried over anhydrous Na₂SO₄, andconcentrated. The resulting residue was purified by columnchromatography to afford the desired product. MS: 160.2 (M+1)⁺.

General Procedures for the Preparation of 2-(3-aminophenyl)propan-2-ol

Step A: Ethyl 3-(dibenzylamino)benzoate 2

To a solution of ethyl 3-aminobenzoate (2 g, 0.012 mmol) and Et₃N (5.26mL, 0.036 mmol) in CH₃CN (30 mL), was added BnBr (4.32 mL, 0.036 mmol)in one portion. The reaction mixture was heated to reflux for 18 hr andthen cooled to room temperature. The mixture was concentrated to drynessin vacuo and the residue was purified by column chromatography(PE:EtOAc=10:1 as eluent) to afford the desired product as a whitesolid. MS: 346.1 (M+1)⁺.

Step B: 2-(3-(dibenzylamino)phenyl)propan-2-ol

To a solution of ethyl 3-(dibenzylamino)benzoate (1.85 g, 5.58 mmol) inanhydrous THF (15 mL) at 0° C. under nitrogen atmosphere was addedMeMgBr (3 M sol. in THF, 5.58 mL, 16.7 mmol) dropwise over 30 min. Thereaction was stirred at room temperature overnight and quenched byaddition of saturated NH₄Cl. The resulting mixture was extracted withethyl acetate (3×50 mL). The combined organic layers were washed withNaHCO₃, water and brine, dried over anhydrous Na₂SO₄, filtered and thenconcentrated to dryness. The residue was purified by columnchromatography (PE:EtOAc=2:1 as eluent) to afford the desired product asa colorless oil. MS: 332.1 (M+1)⁺.

Step C: 2-(3-aminophenyl)propan-2-ol

To a solution of 2-(3-(dibenzylamino)phenyl)propan-2-ol (268 mg, 0.81mmol) in MeOH (5 mL) was added 10% Pd/C (27 mg) in one portion. Thereaction mixture was hydrogenated at room temperature overnight underhydrogen atmosphere. The catalyst was filtered off through Celite andthe filtrate was concentrated to dryness. The residue was purified bycolumn chromatography (PE:EtOAc=1:2 as eluent) to afford the desiredproduct as a yellow solid. MS: 152.1 (M+1)⁺.

General Procedures for the Preparation of2-(3-amino-5-fluoronhenvl)nronan-2-ol

Step A. Methyl 3-fluoro-5-nitrobenzoate

Thionyl chloride (488 mg, 4.1 mmol) was added dropwise to a solution of3-fluoro-5-nitrobenzoic acid (500 mg, 2.7 mmol) in dry methanol (10 mL)at 0° C. under nitrogen atmosphere. The reaction was warmed to roomtemperature and stirred for 6 hr. The reaction mixture was concentratedunder reduced pressure to obtain the corresponding methyl esterhydrochloride as a waxy solid which was used directly in the next step.MS: 200 (M+1)⁺.

Step B. Methyl 3-amino-5-fluorobenzoate

To a solution of methyl 3-fluoro-5-nitrobenzoate (400 mg, 2 mmol) inethanol (10 mL) was added iron powder (560 mg, 10 mmol) and ammoniumchloride (540 mg, 10 mmol) in one portion. The reaction mixture wasstirred at 80° C. for 1 hr. After cooling the reaction, the mixture wasfiltered through Celite. The filtrate was concentrated under reducedpressure to give the desired product. MS: 170 (M+1)⁺.

Step C. Methyl 3-(dibenzylamino)-5-fluorobenzoate

To a solution of methyl 3-amino-5-fluorobenzoate (440 mg, 2.6 mmol) indry DMF (10 mL) was added NaH (187 mg, 7.8 mmol) portionwise, followedby addition of benzyl bromide (1.1 g, 6.5 mmol). The reaction mixturewas stirred at 40° C. for 16 hr and concentrated. The resulting residuewas purified by column chromatography to give the desired product. MS:350 (M+1)⁺.

Step D. 2-(3-(Dibenzylamino)-5-fluorophenyl)propan-2-ol

Methylmagnesium bromide (1 M in THF, 2.4 mL, 2.4 mmol) was dissolved inTHF (5 mL) and placed in an ice-water bath. Methyl3-(dibenzylamino)-5-fluorobenzoate (280 mg, 0.8 mmol) in THF (5 mL) wasthen slowly added to the reaction mixture. This mixture was stirred for3 hr while maintaining an internal temperature range between 15 to 25°C. Then the mixture was cooled to 0° C. and treated with ammoniumchloride solution, then extracted with ethyl acetate (3×30 mL). Thecombined organic layers were dried over anhydrous Na₂SO₄, andconcentrated in vacuo. The residue was purified by column chromatographyon silica gel to give the desired product. MS: 350 (M+1)⁺.

Step E. 2-(3-Amino-5-fluorophenyl)propan-2-ol

To a solution of 2-(3-(dibenzylamino)-5-fluorophenyl)propan-2-ol (150mg, 0.43 mmol) in ethanol (5 mL) was added 10% Pd/C (15 mg) under ahydrogen atmosphere. The reaction mixture was stirred at roomtemperature for 16 hr. The suspension was then filtered through Celiteand the filtrate was concentrated in vacuo. The residue was purified bycolumn chromatography to give the desired product. MS: 170 (M+1)⁺.

General Procedures for the Preparation of ethyl1-(3-aminophenyl)cyclopropanol

Step A. Ethyl 3-(dibenzylamino)benzoate

To a solution of ethyl 3-aminobenzoate (2 g, 0.012 mmol) and Et₃N (5.26mL, 0.036 mmol) in CH₃CN (30 mL) was added BnBr (4.32 mL, 0.036 mmol) inone portion. The reaction mixture was heated to reflux for 18 h andcooled down to room temperature. The mixture was concentrated in vacuoand the resulting residue was purified by column chromatography toafford the desired product as a white solid. MS: 346.1 (M+1)⁺.

Step B. 1-(3-(Dibenzylamino)phenyl)cyclopropanol

To a solution of ethyl 3-(dibenzylamino)benzoate (1.85 g, 5.58 mmol) inanhydrous THF (20 mL) at room temperature under N₂ was added titaniumtetraisopropoxide (0.25 mL, 0.84 mmol) dropwise over 10 min. After onehour of stirring, EtMgBr (THF solution, 4.1 mL, 12.3 mmol) was addeddropwise over 30 min. The reaction mixture was stirred at roomtemperature for 3 h. The resulting mixture was quenched by addition ofsaturated aq. NH₄Cl, and extracted with ethyl acetate (3×50 mL). Thecombined organic layers were washed with NaHCO₃, water and brine, driedover anhydrous Na₂SO₄, and concentrated in vacuo. The residue waspurified by column chromatography (PE:EtOAc=5:1 as eluent) to afford thedesired product as a colorless oil. ¹H NMR (400 MHz, CDCl₃): δ 7.33-7.28(m, 5H), 7.25-7.18 (m, 5H), 7.11 (t, J=8.0 Hz, 1H), 6.80-6.75 (m, 1H),6.61-6.56 (m, 2H), 4.65 (s, 4H), 1.17-1.13 (m, 2H), 0.93-0.90 (m, 2H).MS: 330.1 (M+1)⁺.

Step C. Ethyl 1-(3-aminophenyl)cyclopropanol

To a solution of 1-(3-(dibenzylamino)phenyl)cyclopropanol (1.8 g, 5.45mmol) in MeOH (10 mL) at room temperature was added 10% Pd/C (200 mg) inone portion. The reaction mixture was stirred at room temperature undera hydrogen atmosphere overnight. The suspensino was filtered throughCelite, and the filtrate was concentrated in vacuo. The residue waspurified by column chromatography (PE:EtOAc=2:1 as eluent) to afford thedesired product as a yellow solid. ¹H NMR (400 MHz, CDCl₃): δ 7.10 (t,J=7.8 Hz, 1H), 6.69 (t, J=2.0 Hz, 1H), 6.63-6.60 (m, 1H), 6.56-6.53 (m,1H), 1.22-1.19 (m, 2H), 1.01-0.98 (m, 2H). MS: 150.1 (M+1)⁺.

General Procedures for the Preparation of 3-fluoro-5-(methylthio)aniline

Step A. (3-Fluoro-5-nitrophenyl)(methyl)sulfane

A solution of 3-fluoro-5-nitroaniline (200 mg, 1.28 mmol),1,2-dimethyldisulfane (121 mg, 1.29 mmol) and CH₃CN (3 mL) was stirredat 30° C. Neat isoamyl nitrite (150 mg, 1.28 mmol) was slowly added viasyringe over 5 min. The reaction mixture was slowly heated to refluxover 10 min and maintained at a gentle reflux until N₂ evolution ceased(30-60 min). The reaction mixture was cooled and the solvent was removedin vacuo to afford a dark oil. The resulting oil was purified by columnchromatography to give the desired product as a pale yellow solid.

Step B: 3-Fluoro-5-(methylthio)aniline

To a solution of (3-fluoro-5-nitrophenyl)(methyl)sulfane (90 mg, 0.48mmol) in MeOH (10 mL) was added 10% Pd/C (9 mg) in one portion. Theresulting mixture was purged with H₂ three times and stirred at roomtemperature for 1 h. The suspension was filtered through Celite, and thefilter cake was washed with MeOH (5 mL). The filtrate was concentratedin vacuo to afford the desired product which was used directly in nextstep. MS: 158.0 (M+1)⁺.

General Procedure for the Preparation of(S)-2-oxo-1,3-oxazinane-4-carboxylic acid

To a mixture of (S)-2-amino-4-hydroxybutanoic acid (10 g, 84.0 mmol) and250 mL of aq. NaOH (2 mol/L, 20.4 g, 510 mmol) at 0° C. was added asolution of triphosgene in dioxane (25.3 g in 125 mL dioxane) dropwiseover 1 h. The internal temperature was kept below 5° C. during theaddition. The mixture was then stirred at room temperature for 2 days.The reaction mixture was then concentrated in vacuo, followed byaddition of 200 mL of CH₃CN. The resulting mixture was then heated to60° C. and stirred vigorously for 0.5 h. The hot mixture was filteredimmediately. The filtrate was then concentrated to 100 mL and thedesired product was precipitated out. The crude product was collected byfiltration and used directly in the next step without furtherpurification. MS: 146.0 (M+1)⁺.

General Procedure for the Preparation of(S)-4-(tert-butoxycarbonyl)-6-oxopiperazine-2-carboxylic acid

Step A: (S)-3-Amino-2-(((benzyloxy)carbonyl)amino)propanoic acid

To a mixture of (S)-4-amino-2-(((benzyloxy)carbonyl)amino)-4-oxobutanoicacid (3 g, 11.3 mmol) in MeCN (20 mL), EtOAc (20 mL) and H₂O (10 mL),was added PIAD (4.38 g, 13.5 mmol) in one portion. The reaction mixturewas stirred at room temperature overnight. The resulting mixture wasfiltered, and the filtrate was concentrated in vacuo to afford thedesired product. MS: 239.1 (M+1)⁺.

Step B: (S)-Methyl 3-amino-2-(((benzyloxy)carbonyl)amino)propanoatehydrochloride

To a stirred solution of MeOH (50 mL) was added SOCl₂ (5 mL) dropwise at0° C. The resulting mixture was stirred at 0° C. for 0.5 h before(S)-3-amino-2-(((benzyloxy)carbonyl)amino) propanoic acid (2.6 g, 10mmol) was added. Then the reaction mixture was stirred at roomtemperature overnight and concentrated in vacuo to afford the desiredproduct. MS: 253.1 (M+1)⁺.

Step C: (S)-Methyl3-((2-(benzyloxy)-2-oxoethyl)amino)-2-(((benzyloxy)carbonyl)amino)propaneate

To a solution of (S)-methyl3-amino-2-(((benzyloxy)carbonyl)amino)propanoate hydrochloride (2.6 g,0.01 mol) in THF (40 mL) was added DIPEA (4.0 g, 0.03 mol) at 0° C. Themixture was stirred at 0° C. for 5 min, followed by addition of benzyl2-bromoacetate (4.7 g, 0.02 mol). Then the mixture was allowed to warmto room temperature and stirred overnight. The reaction mixture wasquenched by addition of H₂O and then extracted with EtOAc (3×40 mL). Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄, and concentrated. The resulting residue was purified by columnchromatography to afford the desired product. MS: 401.2 (M+1)⁺.

Step D: (S)-Methyl3-((2-(benzyloxy)-2-oxoethyl)(tert-butoxycarbonyl)amino)-2-(((benzyloxy)carbonyl)amino)propanoate

To a solution of (S)-methyl3-((2-(benzyloxy)-2-oxoethyl)amino)-2-(((benzyloxy)carbonyl)amino)propanoate(3.0 g, 7.5 mmol) in THF (40 mL) was added DIPEA (2.9 g, 22.5 mmol) at0° C. The mixture was stirred at 0° C. for 5 min followed by addition ofdi-tert-butyl dicarbonate (3.27 g, 15 mmol). Then the mixture wasallowed to warm to room temperature and stirred overnight. Afterquenching with a saturated. NaHCO₃ solution, the resulting mixture wasextracted with EtOAc (3×60 mL) and concentrated. The resulting residuewas purified by column chromatography to afford the desired product. MS:501.2 (M+1)⁺.

Step E:(S)-2-((2-Amino-3-methoxy-3-oxopropyl)(tert-butoxycarbonyl)amino)aceticacid

To a solution of (S)-methyl3-((2-(benzyloxy)-2-oxoethyl)(tert-butoxycarbonyl)amino)-2-(((benzyloxy)carbonyl)amino)propanoate(2.5 g, 5 mmol) in MeOH (30 mL) was added 10% Pd/C (250 mg). The mixturewas stirred under hydrogen atmosphere at room temperature overnight. Theresulting suspension was filtered through Celite, and the filtrate wasconcentrated in vacuo to afford the desired product. MS: 277.1 (M+1)⁺.

Step E: (S)-1-tert-Butyl 3-methyl 5-oxopiperazine-1,3-dicarboxylate

To a solution of(S)-2-((2-amino-3-methoxy-3-oxopropyl)(tert-butoxycarbonyl)amino)aceticacid (1.2 g, 4 mmol) in DCM (100 mL) was added DCC (1.34 g, 6 mmol) at5° C. The mixture was stirred at 10° C. for 4 h followed by addition ofEt₃N (0.88 g, 8 mmol). The resulting mixture was stirred at roomtemperature for 18 h and then concentrated. The residue was added toEtOAc (20 mL) and the precipitate was filtered. The filtrate wasconcentrated and the residue was purified by column chromatography toafford the desired product. MS: 259.1 (M+1)⁺.

Step F: (S)-4-(tert-Butoxycarbonyl)-6-oxopiperazine-2-carboxylic acid

To a mixture of (S)-1-tert-butyl 3-methyl5-oxopiperazine-1,3-dicarboxylate (500 mg, 1.9 mmol) in MeOH (20 mL) andTHF (20 mL) was added a solution of LiOH.H₂O (159 mg, 3.8 mmol) in H₂O(10 mL) at 0° C. The mixture was stirred at room temperature for 2 h andthen partitioned between EtOAc (25 mL) and H₂O. The aqueous layer wasacidified with 2N HCl to pH 3-4 and then extracted with EtOAc (3×20 mL).The combined organic layers were washed with brine, dried over anhydrousNa₂SO₄ and concentrated to afford the desired product which was useddirectly in the next reaction. MS: 245.1 (M+1)⁺.

General Procedure for the Preparation of2-bromopyrimidine-4-carbonitrile

Step A: 2-Hydroxy-4-carboxyaldehyde oxime

2-Hydroxy-4-methyl pyrimidine hydrochloride (25.0 g 171 mmol) and sodiumnitrate (17.7 mg, 260 mmol) were slowly added to 200 mL of 50% aceticacid at 0° C. The reaction mixture was stirred at room temperature for 3h. The resulting suspension and the solids were filtered, washed withwater and dried to afford the desired product. ¹H NMR (400 MHz,DMSO-d₆): δ 12.42 (s, 1H), 11.89 (s, 1H), 7.92 (d, J=6.4 Hz, 1H), 7.75(s, 1H), 6.43 (d, J=6.4 Hz, 1H). MS: 140.0 (M+1)⁺.

Step B: 2-Bromopyrimidine-4-carbonitrile

A mixture of 2-hydroxy-4-carboxyaldehyde oxime (9 g, 28.8 mmol),tetrabutyl ammonium bromide (10 g, 71.9 mmol) and phosphorus pentoxide(2 g, 14.4 mmol) in toluene (300 mL) was stirred at 120° C. for 2 h. Theresulting mixture was filtered and the filtrate was concentrated. Theresulting residue was purified by column chromatography to give thedesired compound as a yellow solid. ¹H NMR (400 MHz, CDCl₃): δ 8.82 (d,J=4.8 Hz, 1H), 7.66 (d, J=4.8 Hz, 1H). MS: 185.0 (M+1)⁺.

General Synthetic Procedures for Making Compounds of Formula I:

General Procedures for the UGI Reaction:

A mixture of aldehyde (3.5 mmol) and aniline (3.5 mmol) in MeOH (8 mL)was stirred at room temperature for 30 min. Then the acid (3.5 mmol) wasadded and the reaction mixture was stirred for another 30 min, followedby addition of the isocyanide (3.5 mmol). The resulting mixture was thenstirred at room temperature overnight and quenched with H₂O. Theresulting mixture was partitioned between EtOAc and H₂O. The organiclayer was washed with brine, dried over anhydrous Na₂SO₄, and thenconcentrated. The resulting residue was purified by a standard method toafford the desired product.

General Procedures for the Buchwald Reaction:

A mixture of amine (0.30 mmol), aryl bromide (0.30 mmol), Cs₂CO₃ (129mg, 0.39 mmol), Pd₂(dba)₃ (18 mg, 0.02 mmol) and Xant-Phos (9.4 mg, 0.02mmol) in 1,4-dioxane (10 mL) was stirred under N₂ at 80° C. overnight.After filtration, the filtrate was concentrated in vacuo and the residuewas purified by a standard method to give the desired products.

Example 1 Preparation of (S)-methyl1-methyl-5-oxopyrrolidine-2-carboxylate

Compound 2 was prepared according to the following scheme, using thefollowing protocol.

Step A: (S)-Methyl 1-methyl-5-oxopyrrolidine-2-carboxylate

To a mixture of (S)-5-oxopyrolidine-2-carboxylic acid (5.0 g, 38.8 mmol)in DMF (50 mL) were added anhydrous K₂CO₃ (16 g, 116 mmol) andiodomethane (16.4 g, 116 mmol) at room temperature The resulting mixturewas warmed to 40° C., stirred for 24 h and concentrated in vacuo. Theresidue was precipitated with EtOAc (80 mL) and filtered. The filtercake was washed with EtOAc (2×10 mL). The combined filtrates wereconcentrated and the residue was purified by column chromatography onsilica gel to give the desired product. ¹H-NMR (400 MHz, CDCl₃): δ4.18-4.11 (m, 1H), 3.70 (s, 3H), 2.87 (s, 3H), 2.56-2.29 (m, 3H),2.16-2.04 (m, 1H). MS: 158.1 (M+1)⁺.

Step B: (S)-1-Methyl-5-oxopyrrolidine-2-carboxylic acid

To a solution of (S)-methyl 1-methyl-5-oxopyrrolidine-2-carboxylate (0.6g, 3.8 mmol) in MeOH (6 mL) were added THF (2 mL), H₂O (2 mL) and NaOH(0.45 g, 11.4 mmol) at room temperature The resulting mixture wasstirred at room temperature for 18 h and then acidified with 2 N HCl topH=3-4 at 0° C. The mixture was extracted with EtOAc (3×30 mL), thecombined organic layers were dried over anhydrous Na₂SO₄ andconcentrated to give the crude product as a yellow solid (0.8 g) whichwas used directly in the next step. MS: 142.1 (M−1)⁻.

Step C: Compound 2

2-Chlorobenzaldehyde (117 mg, 0.83 mmol), 3-fluoroaniline (92.5 mg, 0.83mmol), crude (S)-1-methyl-5-oxopyrrolidine-2-carboxylic acid (200 mg,˜60% purity, 0.83 mmol) and 1,1-difluoro-3-isocyanocyclobutane (119 mg,90% purity, 1.0 mmol) were used in the UGI reaction to give the desiredproduct (diastereomeric mixture). ¹H NMR (400 MHz, CDCl₃): δ 8.52 (d,J=4.9 Hz, 0.2H), 8.16 (m, 0.3H), 7.87-7.47 (m, 2H), 7.42-7.31 (m, 1H),7.25-7.11 (m, 2H), 7.08-6.89 (m, 3.3H), 6.74 (d, J=6.0 Hz, 0.7H), 6.57(m, 2H), 4.42-4.26 (m, 1.3H), 4.20-4.08 (m, 0.5H), 4.00 (m, 1H), 3.00(m, 2H), 2.74 (m, 3H), 2.63-1.82 (m, 6H). MS: 494.1 (M+1)⁺.

Example 2 Preparation of(S)—N-(1-(2-chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide

Compounds 3 and 4 were prepared according to the following scheme, usingthe following protocol.

Step A.(S)—N-(1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluoro-phenyl)-5-oxopyrrolidine-2-carboxamide

3-Fluoroaniline (86 mg, 0.78 mmol), 2-chlorobenzaldehyde (109 mg, 0.78mmol), (S)-5-oxopyrrolidine-2-carboxylic acid (100 mg, 0.78 mmol) and1,1-difluoro-4-isocyanocyclohexane (135 mg, 0.91 mmol) were used in theUGI reaction to give the desired product. MS: 508.1 (M+1)⁺.

Step B.(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-1-(pyrimidin-2-yl)pyrrolidine-2-carboxamideand(S)—N—((R)-1-(2-chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-1-(pyrimidin-2-yl)pyrrolidine-2-carboxamide

A mixture of(S)—N-(1-(2-chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(100 mg, 0.20 mmol), 2-bromopyrimidine (47 mg, 0.30 mmol), Cs₂CO₃ (129mg, 0.39 mmol), Pd₂(dba)₃ (18 mg, 0.02 mmol) and Xant-Phos (9.4 mg, 0.02mmol) in 1,4-dioxane (10 mL) was stirred under N₂ at 80° C. overnight.After filtration, the filtrate was concentrated in vacuo and the residuewas purified by a standard method to give the desired products.

(S)—N—((S)-1-(2-chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluoro-phenyl)-5-oxo-1-(pyrimidin-2-yl)pyrrolidine-2-carboxamide.Compound 4 ¹H NMR (400 MHz, CDCl₃): δ 8.71 (d, J=4.8 Hz, 2H), 7.75 (m,1H), 7.33 (m, 2H), 7.18 (m, 1H), 7.09-6.87 (m, 5H), 6.47 (s, 1H), 5.61(d, J=7.6 Hz, 1H), 4.86 (d, J=6.6 Hz, 1H), 3.98 (m, 1H), 3.01-2.84 (m,2H), 2.58 (m, 1H), 2.30-2.20 (m, 1H), 1.93 (m, 7H), 1.47 (m, 2H); MS:586.2 (M+1)⁺.

(S)—N—((R)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-1-(pyrimidin-2-yl)pyrrolidine-2-carboxamide.Compound 3

¹H NMR (400 MHz, CDCl₃): δ 8.75 (dd, J=4.8, 2.0 Hz, 2H), 7.40 (d, J=7.8Hz, 1H), 7.23 (s, 3H), 7.08 (dt, J=11.3, 6.3 Hz, 3H), 6.99 (d, J=3.7 Hz,1H), 6.27 (s, 1H), 6.13-5.92 (m, 1H), 5.02 (m, 1H), 4.76 (m, 1H), 3.92(m, 1H), 2.88 (m, 1H), 2.67-2.46 (m, 1H), 2.44-2.19 (m, 2H), 2.00 (m,8H). MS: 586.1 (M+1)⁺.

The following analogs were synthesized via the procedures set forthabove, using the appropriate aldehyde, amine, carboxylic acid,isocyanide and halo-substituted-aromatic ring or heteroaromatic ringusing the reagents and solvents set forth above or similar reagents andsolvents thereof, and purified via standard methods.

¹H NMR (400 MHz, CDCl₃): δ 8.75 (d, J=4.8 Hz, 2H), 7.35 (m, 3H),7.25-6.81 (m, 5H), 6.28 (s, 1H), 5.84 (d, J=7.5 Hz, 1H), 4.76 (m, 1H),3.98-3.59 (m, 1H), 2.92 (m, 1H), 2.58 (m, 1H), 2.35-2.20 (m, 1H), 2.07(m, 1H), 1.83 (m, 2H), 1.57 (m, 4H), 1.46-1.17 (m, 4H). MS: 550.2(M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.73 (m, 2H), 7.80 (s, 1H), 7.35 (s, 1H),7.23-6.72 (m, 6H), 6.47 (s, 1H), 5.49 (d, J=7.7 Hz, 1H), 4.87 (d, J=6.6Hz, 1H), 4.74-4.42 (m, 1H), 3.86 (d, J=8.0 Hz, 1H), 3.19-2.77 (m, 1H),2.56 (m, 1H), 2.44-2.21 (m, 1H), 2.13-1.73 (m, 4H), 1.60 (s, 2H), 1.26(m, 4H). MS: 550.2 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.69 (s, 2H), 7.76 (s, 1H), 7.49-6.68 (m,7H), 6.44 (s, 1H), 6.19 (s, 1H), 4.93 (m, 3H), 2.23 (m, 8H). MS: 540.1(M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.81 (d, J=4.9 Hz, 1H), 8.66 (d, J=2.7 Hz,1H), 8.04-7.79 (m, 1H), 7.49-7.31 (m, 1H), 7.13-6.92 (m, 6H), 6.60 (m,1H), 6.25-5.95 (m, 1H), 5.68 (m, 1H), 4.73 (dd, J=16.0, 6.9 Hz, 1H),4.39 (m, 1H), 2.98 (m, 3H), 2.53 (m, 4H), 2.14-1.93 (m, 1H). MS: 592.1(M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.46-8.32 (m, 1.7H), 7.78-7.61 (m, 1.5H),7.39 (m, 1.5H), 7.23 (m, 1.6H), 7.13-6.88 (m, 4H), 6.40 (m, 1H), 6.11(m, 1H), 5.01-4.77 (m, 1H), 4.26 (m, 1H), 3.51 (d, J=5.5 Hz, 0.3H),3.13-2.75 (m, 3H), 2.61-2.22 (m, 3H), 2.17-1.90 (m, 1H). MS: 557.1(M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.56 (m, 2H), 8.16 (s, 1.3H), 7.74 (s, 1H),7.36 (s, 2.6H), 7.19 (s, 1H), 7.12-6.82 (m, 3H), 6.52 (m, 2H), 6.19 (m,1H), 4.65-4.48 (m, 1H), 4.26 (m, 1.3H), 3.90-3.82 (m, 0.3H), 2.87 (m,3H), 2.64-1.98 (m, 6H). MS: 557.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 7.98 (m, 1H), 7.65 (m, 2H), 7.44-7.30 (m,2H), 7.03 (m, 6H), 6.51 (m, 1H), 6.36 (s, 1H), 5.12 (d, J=6.3 Hz, 1H),4.33 (s, 1H), 3.97 (s, 3H), 3.10-2.63 (m, 3H), 2.60-2.00 (m, 5H). MS:587.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.32 (m, 1H), 8.05 (t, J=8.6 Hz, 1H), 7.69(s, 1H), 7.45-7.30 (m, 1H), 7.25-6.78 (m, 6H), 6.38 (m, 2H), 4.88 (m,1H), 4.33 (s, 1H), 3.89 (s, 3H), 3.11-2.72 (m, 3H), 2.66-2.29 (m, 3H),2.23-1.86 (m, 2H). MS: 587.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 7.93 (m, 1H), 7.56 (m, 2H), 7.21 (m, 3H),7.10-6.87 (m, 3H), 6.42 (m, 3H), 5.04 (m, 1H), 4.25 (m, 1H), 3.97 (d,J=6.1 Hz, 3H), 3.10-2.69 (m, 3H), 2.60-2.15 (m, 4H), 2.12-1.87 (m, 1H).MS: 587.2 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.19 (m, 1H), 7.79-7.33 (m, 3H), 7.28-7.06(m, 4H), 7.06-6.83 (m, 4H), 6.47-6.32 (m, 2H), 5.09-4.91 (m, 1H), 4.25(m, 1H), 3.09-2.60 (m, 4H), 2.57 (s, 3H), 2.53-1.99 (m, 5H). MS: 571.0(M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.26 (d, J=8.5 Hz, 1H), 8.15 (s, 1H), 7.64(s, 1H), 7.48 (m, 1H), 7.32 (d, J=7.5 Hz, 1H), 7.14 (m, 2H), 7.04-6.83(m, 3H), 6.40 (s, 1H), 6.04 (s, 1H), 4.89 (m, 1H), 4.31 (s, 1H), 2.89(m, 3H), 2.48 (m, 2H), 2.40-2.27 (m, 3H), 2.26-1.84 (m, 3H). MS: 571.2(M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.30-8.15 (m, 2H), 7.68 (s, 1H), 7.38 (m,1H), 7.24-6.85 (m, 6H), 6.46-6.16 (m, 2H), 4.94 (d, J=6.0 Hz, 1H), 4.32(s, 1H), 3.10-2.74 (m, 3H), 2.60-2.43 (m, 2H), 2.36 (m, 4H), 2.23-1.91(m, 2H). MS: 571.2 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.17 (d, J=8.3 Hz, 1H), 7.56 (m, 2H),7.25-6.96 (m, 5H), 6.89 (m, 2H), 6.42 (s, 1H), 6.21 (s, 1H), 5.12-4.96(m, 1H), 4.31 (m, 1H), 3.14-2.74 (m, 3H), 2.55 (s, 3H), 2.51-2.28 (m,3H), 2.20 (m, 1H), 2.05-1.87 (m, 1H). MS: 571.2 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.72 (m, 1H), 7.88 (m, 1H), 7.65 (s, 1H),7.57-7.30 (m, 2H), 7.23-7.09 (m, 2H), 7.02 (s, 2H), 6.96-6.83 (m, 1H),6.44 (s, 1H), 6.05 (d, J=6.5 Hz, 1H), 5.31-4.93 (m, 1H), 4.33 (s, 1H),3.02 (m, 2H), 2.86 (m, 1H), 2.63-2.45 (m, 2H), 2.44-2.23 (m, 2H), 2.01(m, 1H). MS: 625.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.91-8.34 (m, 2H), 8.03 (s, 1H), 7.79-7.34(m, 3H), 7.22-6.75 (m, 5H), 6.46 (s, 1H), 6.02 (d, J=6.5 Hz, 1H), 4.95(dd, J=9.4, 3.1 Hz, 1H), 4.35 (s, 1H), 3.13-2.76 (m, 3H), 2.68-1.83 (m,5H). MS: 625.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.65 (d, J=23.6 Hz, 2H), 7.87 (s, 1H),7.59-7.29 (m, 3H), 7.26-6.71 (m, 5H), 6.59 (s, 1H), 6.28 (s, 1H), 4.83(d, J=8.2 Hz, 1H), 4.12 (s, 1H), 3.10-2.62 (m, 3H), 2.56 (m, 1H),2.36-1.84 (m, 4H). MS: 625.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.74 (s, 1H), 8.53 (s, 1H), 7.71 (s, 1H),7.31 (d, J=8.3 Hz, 1H), 7.25-6.80 (m, 6H), 6.44 (s, 1H), 6.08 (s, 1H),4.95 (m, 1H), 4.35 (s, 1H), 3.15-2.76 (m, 3H), 2.66-2.17 (m, 4H), 2.03(s, 1H). MS: 625.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.29 (dd, J=8.1, 2.0 Hz, 1H), 7.74 (m, 2H),7.31 (m, 2H), 7.22-7.12 (m, 2H), 7.00 (s, 2H), 6.93 (m, 1H), 6.67 (dd,J=7.9, 2.4 Hz, 1H), 6.46 (m, 1H), 6.06 (m, 1H), 4.86 (m, 1H), 4.35 (m,1H), 2.93 (m, 3H), 2.59-2.39 (m, 2H), 2.23 (m, 1H), 2.02 (m, 1H). MS:575.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.40 (m, 1H), 8.24 (m, 1H), 7.71 (d, J=7.7Hz, 1H), 7.49-7.30 (m, 2H), 7.28-7.21 (m, 1H), 7.12 (m, 2H), 7.04-6.88(m, 3H), 6.67 (m, 1H), 6.42 (s, 2H), 4.90 (m, 1H), 4.27 (m, 1H),3.07-2.76 (m, 3H), 2.58-2.29 (m, 3H). MS: 575.0 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.27 (m, 1H), 7.64-7.30 (m, 3H), 7.27-6.62(m, 7H), 6.47-6.30 (m, 1H), 6.28-6.07 (m, 1H), 5.00-4.55 (m, 1H), 4.26(m, 1H), 3.12-2.67 (m, 3H), 2.65-2.36 (m, 3H), 2.22 (m, 2H). MS: 575.1(M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.37 (t, J=8.9 Hz, 1H), 7.63 (m, 2H),7.49-6.84 (m, 8H), 6.44 (s, 1H), 5.94 (m, 1H), 5.07-4.74 (m, 1H), 4.25(d, J=51.6 Hz, 1H), 3.10-2.67 (m, 3H), 2.63-1.85 (m, 5H), 1.25 (s, 1H).MS: 591.1 (M+1)⁺.

¹H NMR (400 MHz, DMSO-d₆): δ 8.38 (d, J=8.2 Hz, 1H), 7.86-7.34 (m, 4H),7.25-6.79 (m, 6H), 6.46 (s, 1H), 5.99 (s, 1H), 4.95 (d, J=9.2 Hz, 1H),4.34 (s, 1H), 3.12-2.70 (m, 3H), 2.63-1.87 (m, 6H). MS: 591.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.59-8.19 (m, 2H), 7.82-7.57 (m, 2H),7.45-7.34 (m, 2H), 7.01 (m, 4H), 6.45 (s, 1H), 5.94 (s, 1H), 4.89 (dd,J=9.3, 3.1 Hz, 1H), 4.30 (m, 1H), 3.21-2.69 (m, 3H), 2.61-1.88 (m, 5H).MS: 591.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.63-8.03 (m, 2H), 7.67 (s, 1H), 7.23-6.65(m, 8H), 6.45-5.93 (m, 2H), 4.84 (m, 1H), 4.23 (m, 1H), 3.04-2.65 (m,4H), 2.65-1.83 (m, 5H). MS: 591.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.79-8.51 (m, 2H), 7.88 (s, 1H), 7.51-7.29(m, 2H), 7.22 (m, 2H), 7.08 (t, J=7.3 Hz, 1H), 6.99 (t, J=7.2 Hz, 1H),6.78 (s, 1H), 6.51 (d, J=5.8 Hz, 1H), 6.28 (s, 1H), 4.79 (m, 1H), 4.14(s, 1H), 3.02-2.66 (m, 3H), 2.55 (m, 1H), 2.33-1.99 (m, 4H). MS: 582.1(M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.74 (s, 1H), 8.52 (s, 1H), 7.85-7.30 (m,3H), 7.24-6.79 (m, 5H), 6.43 (s, 1H), 6.12 (s, 1H), 4.92 (d, J=6.8 Hz,1H), 4.34 (s, 1H), 2.90 (m, 3H), 2.64-2.46 (m, 1H), 2.46-2.11 (m, 3H),1.97 (m, 1H). MS: 582.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.66-8.38 (m, 2H), 7.90 (d, J=7.0 Hz, 1H),7.68 (s, 1H), 7.37 (m, 1H), 7.25-6.80 (m, 6H), 6.44 (s, 1H), 5.97 (d,J=6.6 Hz, 1H), 4.91 (d, J=6.7 Hz, 1H), 4.32 (s, 1H), 3.30-2.78 (m, 4H),2.41 (m, 4H), 2.02 (s, 1H). MS: 582.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.58 (d, J=9.3 Hz, 1H), 8.11 (d, J=8.7 Hz,1H), 7.97 (d, J=8.5 Hz, 1H), 7.86-7.59 (m, 3H), 7.48 (m, 2H), 7.18 (m,3H), 6.97 (m, 3H), 6.38 (s, 1H), 6.11 (s, 1H), 5.20 (s, 1H), 4.30 (s,1H), 3.09-2.77 (m, 3H), 2.67-2.44 (m, 2H), 2.36-2.21 (m, 2H), 2.10-1.92(m, 1H). MS: 607.2 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 8.69 (d, J=4.8 Hz, 2H), 7.71 (s, 1H), 7.31(m, 1H), 7.18 (m, 1H), 7.13-6.77 (m, 6H), 6.46 (s, 1H), 6.22 (s, 1H),5.00-4.62 (m, 1H), 4.35 (s, 1H), 3.19-2.71 (m, 3H), 2.69-1.83 (m, 5H).MS: 451.2 (M+1)⁺.

¹H NMR (400 MHz, DMSO-d₆): δ 8.15-8.01 (m, 1H), 7.62-7.52 (m, 1H),7.31-6.69 (m, 9H), 6.24 (s, 1H), 5.65-4.66 (m, 1H), 2.60 (m, 1H),2.20-2.05 (m, 3H), 1.76-0.83 (m, 4H). MS: 451.2 (M+1)⁺.

¹H NMR (400 MHz, DMSO-d₆): δ 9.70 (s, 1H), 8.48-8.26 (m, 2H), 7.72 (s,1H), 7.46-7.31 (m, 1H), 7.28-7.15 (m, 2H), 7.13-6.89 (m, 3H), 6.55-6.14(m, 2H), 4.82 (m, 1H), 4.26 (m, 1H), 2.90 (m, 3H), 2.64-2.40 (m, 2H),2.34-1.99 (m, 3H). MS: 558.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 7.54 (d, J=3.5 Hz, 1H), 7.45-7.29 (m, 3H),7.28-6.95 (m, 6H), 6.44 (d, J=6.0 Hz, 1H), 6.24 (s, 1H), 4.92 (m, 1H),4.25 (s, 1H), 3.11-2.79 (m, 3H), 2.61 (m, 1H), 2.43 (m, 1H), 2.39-2.27(m, 2H), 2.27-2.11 (m, 1H). MS: 563.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 7.66 (s, 1H), 7.48 (s, 1H), 7.35 (s, 1H),7.26-6.82 (m, 8H), 6.43 (s, 1H), 6.09 (d, J=6.3 Hz, 1H), 4.98 (d, J=8.7Hz, 1H), 4.34 (s, 1H), 3.08-2.84 (m, 2H), 2.63-2.36 (m, 4H), 2.32 (m,1H), 2.15 (m, 1H). MS: 563.1 (M+1)⁺.

¹H NMR (400 MHz, CDCl₃): δ 7.78-7.49 (m, 2H), 7.39 (m, 4H), 7.24-6.82(m, 4H), 6.38 (m, 3H), 5.94 (m, 1H), 4.50 (m, 1H), 4.22 (m, 1H),3.10-2.59 (m, 3H), 2.59-1.99 (m, 6H). MS: 556.2 (M+1)⁺.

Example 3 Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-1-(thiazol-4-yl)pyrrolidine-2-carboxamide

Compounds 42 and 43 were prepared according to the following scheme,using the following protocol.

A mixture(2S)—N-(1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(200 mg, 0.417 mmol), 4-bromothiazole (0.045 mL, 0.626 mmol, 1.5 eq),K₃PO₄ (124 mg, 0.585 mmol, 1.4 eq), CuI (8 mg, 0.1 eq) andtrans-1,2-diaminocyclohexane (0.24 eq) in dioxane (2 mL) was stirred at110° C. under microwave for 30 min. The resulting mixture was filteredthrough a Celite pad. The filtrate was concentrated and the residue waspurified by a standard method to give the desired product.

(S)—N—((R)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-1-(thiazol-4-yppyrrolidine-2-carboxamide(Compound 42)

¹H NMR (400 MHz, CDCl₃): δ 8.68 (d, J=2.1 Hz, 1H), 7.65 (m, 5H),7.30-6.90 (m, 4H), 6.47 (s, 1H), 6.23 (s, 1H), 4.88 (dd, J=9.3, 3.0 Hz,1H), 4.20 (s, 1H), 3.17-2.63 (m, 3H), 2.58-1.99 (m, 5H). MS: 563.1(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-1-(thiazol-4-yl)pyrrolidine-2-carboxamide(Compound 43)

¹H NMR (400 MHz, CDCl₃): δ 8.60 (s, 1H), 8.06-7.56 (m, 2H), 7.35 (s,1H), 7.22-6.79 (m, 5H), 6.42 (s, 1H), 6.13 (s, 1H), 4.96 (d, J=7.8 Hz,1H), 4.25 (m, 1H), 3.14-2.70 (m, 4H), 2.63-2.21 (m, 4H). MS: 563.1(M+1)⁺.

Example 4 Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-1-(pyridin-2-ylmethyl)pyrrolidine-2-carboxamide

Compound 44 was prepared according to the following scheme, using thefollowing protocol.

Compound 44

To a solution of(2S)—N-(1-(2-chlorophenyl)-2-(3,3-difluorocyclobutyl-amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(200 mg, 0.42 mmol) in dry DMF (20 mL) was added NaH (20 mg, 0.84 mmol)at 0° C. The mixture was stirred at this 0° C. for 0.5 h followed byaddition of 2-(bromomethyl)pyridine (106 mg, 0.42 mmol). The mixture wasthen allowed warm to room temperature and stirred overnight. Theresulting mixture was slowly added dropwise to 100 mL of water, and thenextracted with EtOAc (3×20 mL). The combined organic layers were washedwith saturated aq. LiCl, dried over anhydrous Na₂SO₄ and concentrated invacuo. The residue was purified by a standard method to afford thedesired product. ¹H NMR (400 MHz, CDCl₃): δ 8.51 (s, 1H), 7.88-7.37 (m,3H), 7.19-5.95 (m, 10H), 5.14 (m, 1H), 4.34 (m, 1H), 4.10 (m, 2H), 3.00(m, 2H), 2.81-1.57 (m, 6H). MS: 571.2 (M+1)⁺.

Example 5 Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-3-hydroxy-2-(pyrimidin-2-ylamino)propanamide

Compound 9 was prepared according to the following scheme, using thefollowing protocol.

Step A: (S)-2-Oxooxazolidine-4-carboxylic acid

To a solution of NaOH (0.8 g, 20 mmol) in water (4 mL) was added(S)-2-(benzyloxycarbonylamino)-3-hydroxypropanoic acid (1 g, 4.2 mmol)portionwise at 0° C. over 3 min. The resulting solution was warmed tor.t and stirred for 2 h. After cooling to 0° C., the solution wasadjusted to pH=1-2 with 2 N HCl. The mixture was extracted with EtOAc(4×10 mL). The combined organic layers were dried over anhydrous Na₂SO₄and concentrated in vacuo to give the desired product as a white solid.¹H NMR (400 MHz, DMSO-d₆): δ 13.93-12.30 (m, 1H), 8.15 (s, 1H), 4.49 (t,J=8.6 Hz, 1H), 4.32 (m, 2H); MS: 130.0 (M−1)⁻.

Step B:(4S)—N-(1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluoro-phenyl)-2-oxooxazolidine-4-carboxamide

2-Chlorobenzaldehyde (160 mg, 1.14 mmol), 3-fluoroaniline (127 mg, 1.14mmol), (S)-2-oxooxazolidine-4-carboxylic acid (150 mg, 1.14 mmol) and1,1-difluoro-3-isocyanocyclobutane (181 mg, 90% of purity, 1.37 mmol)were used in the UGI reaction to give the desired product as a whitesolid. ¹H NMR (400 MHz, CDCl₃): δ 8.15-8.01 (m, 1H), 7.62-7.52 (m, 1H),7.31-6.69 (m, 9H), 6.24 (s, 1H), 5.65-4.66 (m, 4H), 2.60 (m, 1H),2.20-2.05 (m, 3H), 1.76-1.51 (m, 5H), 1.29-0.83 (m, 5H); MS: 482.1(M+1)+.

Step C:(S)—N—((R)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-2-oxo-3-(pyrimidin-2-yl)oxazolidine-4-carboxamideand(S)—N—((S)-1-(2-chloro-phenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-2-oxo-3-(pyrimidin-2-yl)oxazolidine-4-carboxamide

A mixture of(4S)—N-(1-(2-chlorophenyl)-2-(3,3-difluorocyclo-butylamino)-2-oxoethyl)-N-(3-fluorophenyl)-2-oxooxazolidine-4-carboxamide(350 mg, 0.73 mmol), 2-bromopyrimidine (150 mg, 0.94 mmol), Cs₂CO₃ (500mg, 1.52 mmol), Pd₂(dba)₃ (66 mg, 0.07 mmol) and Xant-Phos (42 mg, 0.07mmol) in 1,4-dioxane (15 mL) was stirred under N₂ at 80° C. for 18 h andthen filtered through a Celite pad. The filtrate was concentrated invacuo and the residue was purified a standard method to give(S)—N—((R)-1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-2-oxo-3-(pyrimidin-2-yl)oxazolidine-4-carboxamide(8). ¹H NMR (400 MHz, CDCl₃): δ 8.73 (d, J=4.8 Hz, 2H), 7.95 (s, 0.8H),7.74 (s, 0.2H), 7.41 (d, J=7.5 Hz, 1.6H), 7.24 (t, J=7.2 Hz, 1H),7.17-6.94 (m, 4.3H), 6.73 (d, J=6.7 Hz, 1H), 6.48 (d, J=73.8 Hz, 2H),4.93 (s, 1H), 4.41 (dd, J=8.6, 4.8 Hz, 1H), 4.29 (t, J=8.6 Hz, 1H), 4.14(m, 1H), 2.80 (m, 2H), 2.21 (s, 1H), 2.18-2.07 (m, 1H); MS: 560.1(M+1)⁺, and(S)—N—((S)-1-(2-chloro-phenyl)-2-(3,3-difluorocyclo-butylamino)-2-oxoethyl)-N-(3fluoro-phenyl)-2-oxo-3-(pyrimidin-2-yl)oxazolidine-4-carboxamide(9). ¹H NMR (400 MHz, CDCl₃): δ 8.68 (d, J=4.8 Hz, 2H), 7.65 (s, 1H),7.30 (s, 1H), 7.18 (s, 1H), 7.13-6.86 (m, 5H), 6.50 (s, 1H), 6.38 (m,1H), 5.00 (m, 1H), 4.43 (dd, J=8.7, 4.8 Hz, 1H), 4.32 (m, 1H), 4.20 (m,1H), 2.99 (m, 2H), 2.50 (m, 2H). MS: 560.1 (M+1)⁺.

Example 6 Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)-amino)-2-oxoethyl)-N-(3-fluorophenyl)-6-oxo-1-(pyrimidin-2-yl)piperidine-2-carboxamide

Compounds 19 and 20 were prepared according to the following scheme,using the following protocol.

Step A. (S)-6-Oxopiperidine-2-carboxylic acid

A solution of (S)-2-aminohexanedioic acid (470 mg, 2.9 mmol) in 20% AcOH(5 mL) was stirred at 110° C. overnight. The solvent was removed invacuo and the residue was dissolved in EtOH (10 mL). The unreacted aminoacid was precipitated and filtered off. The filtrate was concentrated togive the crude desired product which was used directly in the next step.MS: 142.1 (M−1)⁻.

Step B.(S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluoro-phenyl)-6-oxopiperidine-2-carboxamide

3-Fluoroaniline (217 mg, 1.96 mmol), 2-chlorobenzaldehyde (274 mg, 1.96mmol), (S)-6-oxopiperidine-2-carboxylic acid (280 mg, 1.96 mmol) and1,1-difluoro-3-isocyanocyclobutane (280 mg, 1.96 mmol) were used in theUGI reaction to give the desired product. MS: 494.1 (M+1)⁺.

Step C.(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-6-oxo-1-(pyrimidin-2-yl)piperidine-2-carboxamideand(S)—N—((R)-1-(2-chloro-phenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-6-oxo-1-(pyrimidin-2-yl)piperidine-2-carboxamide

A mixture consisting of(1R)—N-(1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-3-oxo-2-(pyrimidin-2-yl)cyclohexanecarboxamide(250 mg, 0.51 mmol), 2-bromopyrimidine (121 mg, 0.76 mmol), Cs₂CO₃ (331mg, 1.01 mmol), Pd₂(dba)₃ (46 mg, 0.05 mmol) and Xant-Phos (29 mg, 0.04mmol) in 1,4-dioxane (15 mL) was stirred under N₂ at 80° C. overnightand then filtered. The filtrate was concentrated in vacuo and theresidue was purified by a standard method to give the desired products.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluoro-phenyl)-6-oxo-1-(pyrimidin-2-yl)piperidine-2-carboxamide(Compound 19). ¹H NMR (400 MHz, CDCl₃): δ 8.73 (m, 2H), 7.70 (s, 1H),7.26-6.95 (m, 6H), 6.87 (t, J=7.2 Hz, 1H), 6.53 (s, 1H), 6.33 (s, 1H),4.77 (d, J=5.3 Hz, 1H), 4.33 (s, 1H), 3.01 (d, J=5.5 Hz, 2H), 2.85-2.28(m, 4H), 2.05 (m, 2H), 1.81 (s, 2H). MS: 571.1 (M+1)⁺.

(S)—N—((R)-1-(2-Chloro-phenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluoro-phenyl)-6-oxo-1-(pyrimidin-2-yl)piperidine-2-carboxamide(Compound 20)

¹H NMR (400 MHz, CDCl₃): δ 8.74 (d, J=4.8 Hz, 2H), 7.99 (m, 1H),7.56-7.32 (m, 1H), 7.27-6.85 (m, 6H), 6.72 (s, 1H), 6.51 (m, 1H),4.67-4.48 (m, 1H), 4.34-4.01 (m, 1H), 2.95-2.60 (m, 2H), 2.59-2.40 (m,1H), 2.40-2.19 (m, 2H), 2.15-2.00 (m, 2H), 1.97-1.59 (m, 4H). MS: 571.1(M+1)⁺.

Example 7 Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-4-(pyrimidin-2-yl)morpholine-3-carboxamide

Compound 30 was prepared according to the following scheme, using thefollowing protocol.

Step A: (S)-3-Hydroxy-2-(4-methoxybenzylamino)propanoic acid

(S)-2-amino-3-hydroxy-propanoic acid (8.4 g, 80 mmol) was dissolved in asolution of NaOH (3.2 g, 80 mmol) in H₂O (40 mL). After cooling to 10°C., 4-methoxybenzaldehyde (21.7 g, 160 mmol) was added dropwise over 10min. The mixture was stirred at room temperature for 30 min and thencooled to 0° C. NaBH₄ (1.67 g, 44 mmol) was added portionwise and theresulting mixture was warmed slowly to r.t and stirred for 2 h. Themixture was washed with Et₂O (2×50 mL). The aqueous phase was adjustedto pH 4.5 with 2 N HCl at 0° C. The precipitate was filtered, washedwith petroleum ether (20 mL) and dried in vacuo to give the desiredproduct as a white solid. MS: 226.1 (M+1)⁺.

Step B: (S)-Benzyl4-(4-methoxybenzyl)-5-oxomorpholine-3-carboxylate

(S)-3-Hydroxy-2-((4-methoxybenzyl)amino)propanoic acid (5.0 g, 22 mmol)was dissolved in a solution of NaOH (1.15 g, 29 mmol) in H₂O (60 mL).After cooling to 0° C., 2-chloroacetyl chloride (3.6 mL, 44 mmol) wasadded dropwise followed by aq. NaOH (30% wt) to keep pH=13. Afterstirring for another 4 h, the reaction was cooled to 0° C. and acidifiedwith 2 N HCl to adjust pH=2-3. The resulting mixture was extracted withEtOAc (2×30 mL). The combined organic layers were dried over anhydrousNa₂SO₄ and concentrated. The residue was dissolved in acetone (150 mL)and then treated with BnBr (9.7 g, 51 mmol) and DIPEA (19 mL, 111 mmol).The reaction mixture was stirred for 24 h at room temperature andconcentrated in vacuo. The residue was purified by column chromatographyto afford the desired product as a white solid. MS: 356.1 (M+1)⁺.

Step C: (S)-Benzyl5-oxomorpholine-3-carboxylate

To a solution of (S)-benzyl4-(4-methoxybenzyl)-5-oxomorpholine-3-carboxylate (200 mg, 0.56 mmol) inCH₃CN (5 mL) and H₂O (5 mL) was added CAN (ceric ammonium nitrate) (1.5g, 2.8 mmol) at 0° C. The resulting mixture was stirred at 0° C. for 1h. DIPEA was added at 0° C. to adjust the pH to 6-7 and the mixture wasconcentrated in vacuo. The residue was purified by column chromatographyto afford the desired product as a white solid. MS: 236.1 (M+1)⁺.

Step D: (S)-5-Oxomorpholine-3-carboxylic acid

To a mixture of (S)-benzyl 5-oxomorpholine-3-carboxylate (160 mg, 0.7mmol) in MeOH (8 mL) was added 10% Pd/C (about 5 mg). The reaction wasstirred under an atmosphere of hydrogen for 30 min at room temperature.The reaction mixture was filtered through a Celite pad and concentratedin vacuo to afford the desired product as a white solid. MS: 146.1(M+1)⁺.

Step E:(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxomorpholine-3-carboxamide

3-Chlorobenzaldehyde (104 mg, 0.74 mmol), 3-fluoroaniline (83 mg, 0.74mmol), (S)-5-oxomorpholine-3-carboxylic acid (108 mg, 0.74 mmol) and1,1-difluoro-3-isocyanocyclobutane (248 mg, 1.48 mmol) were used in theUGI reaction to afford the desired product. MS: 496.1 (M+1)⁺.

Step F: Compound 30

A mixture of(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluoro-cyclobutyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxomorpholine-3-carboxamide(100 mg, 0.2 mmol), 2-bromopyrimidine (36 mg, 0.22 mmol), Pd₂(dba)₃ (28mg, 0.03 mmol), XantPhos (16 mg, 0.03 mmol) and Cs₂CO₃ (160 mg, 0.5mmol) in 1,4-dioxane (4 mL) was stirred at 100° C. for 3.5 h under N₂.The reaction mixture was then cooled to room temperature and filtered.The solid was washed with DCM (2×20 mL). The filtrate was evaporated andthe residue was purified by a standard method to afford the desiredproduct. ¹H NMR (400 MHz, CDCl₃): δ 8.77 (m, 2H), 7.85 (m, 1H), 7.41 (s,1H), 7.28-7.21 (m, 1H), 7.21-7.10 (m, 2H), 7.09-6.90 (m, 3H), 6.87 (m,1H), 6.68-6.33 (m, 2H), 4.80 (m, 1H), 4.43-4.22 (m, 2H), 4.13 (m, 2H),3.94 (m, 1H), 2.99 (m, 1H), 2.86 (m, 1H), 2.63-2.26 (m, 2H). MS: 474.1(M+1)⁺.

Example 8

The following analogs were synthesized via the procedure set forthabove, using the appropriate aldehyde, amine, carboxylic acid,isocyanide and halo-substituted aromatic ring or heterocyclic(heteroaromatic) ring using the reagents and solvents set forth above,and purified via standard methods.

(2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclopentypamino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-1-(pyrimidin-2-yl)pyrrolidine-2-carboxamide(Racemic)—Compound 73

¹H NMR (400 MHz, CDCl₃): δ 8.71 (d, J=4.8 Hz, 2H), 7.72 (s, 1H), 7.37(s, 1H), 7.18 (s, 1H), 7.11-6.85 (m, 5H), 6.47 (s, 1H), 5.70 (d, J=7.3Hz, 1H), 4.86 (d, J=7.0 Hz, 1H), 4.53 (d, J=6.3 Hz, 1H), 3.51 (s, 1H),2.95-2.88 (m, 1H), 2.64-2.47 (m, 2H), 2.40-1.65 (m, 8H). MS: 572.2(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 64

¹H NMR (400 MHz, CDCl₃): δ 8.74 (s, 1H), 8.52 (s, 1H), 7.72 (d, J=7.1Hz, 1H), 7.43-7.33 (m, 1H), 7.25-7.17 (m, 1H), 7.13-6.81 (m, 4H), 6.43(s, 1H), 6.12 (s, 1H), 4.92 (d, J=6.8 Hz, 1H), 4.37-4.28 (m, 1H),3.10-2.82 (m, 3H), 2.59-2.49 (m, 2H), 2.42-2.36 (m, 1H), 2.31-2.22 (m,1H), 2.06-1.88 (m, 2H). MS: 582.1 (M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-2-((3,3-difluorocyclobutyl)amino)-2-oxo-1-phenylethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 138

¹H NMR (400 MHz, CDCl₃): δ 8.78 (s, 1H), 8.44 (d, J=4.9 Hz, 1H), 7.65(s, 1H), 7.39-7.15 (m, 6H), 7.14-6.92 (m, 4H), 6.65 (m, 1H), 6.16 (s,1H), 5.82 (s, 1H), 4.86 (d, J=6.8 Hz, 1H), 4.31 (s, 1H), 3.15-2.77 (m,3H), 2.68-1.91 (m, 5H). MS: 548.2 (M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-2-((3,3-difluorocyclobutyl)amino)-1-(2-fluorophenyl)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 149

¹H NMR (400 MHz, CDCl₃): δ 8.74 (m, 1H), 8.50 (d, J=4.2 Hz, 1H), 7.65(s, 1H), 7.45-7.14 (m, 4H), 7.13-6.69 (m, 5H), 6.25 (m, 2H), 4.88 (dd,J=9.2, 3.1 Hz, 1H), 4.33 (s, 1H), 3.21-2.72 (m, 3H), 2.65-1.88 (m, 5H).MS: 566.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 68

¹H NMR (400 MHz, CDCl₃): δ 8.95 (d, J=4.7 Hz, 1H), 7.68 (s, 1H), 7.34(d, J=4.6 Hz, 2H), 7.16 (s, 1H), 7.04 (d, J=3.6 Hz, 3H), 6.92 (s, 2H),6.51 (s, 1H), 5.92 (s, 1H), 4.81 (d, J=9.5 Hz, 1H), 4.33 (s, 1H), 2.91(m, 3H), 2.64-2.26 (m, 4H), 2.01 (s, 1H). MS: 583.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 85

¹H NMR (400 MHz, CDCl₃): δ 8.98 (d, J=4.7 Hz, 1H), 7.74 (s, 1H), 7.38(dd, J=11.2, 5.7 Hz, 2H), 7.06 (m, 5H), 6.52 (s, 1H), 5.47 (d, J=7.7 Hz,1H), 4.85 (d, J=9.2 Hz, 1H), 3.99 (s, 1H), 2.93 (dd, J=18.6, 8.9 Hz,1H), 2.62 (d, J=9.5 Hz, 1H), 2.36 (s, 1H), 1.97 (m, 7H), 1.57-1.38 (m,2H). MS: 611.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3,5-difluoro-phenyl)-5-oxo-1-(pyrimidin-2-yl)pyrrolidine-2-carboxamide(Single Enantiomer)—Compound 70

¹H NMR (400 MHz, CDCl₃): δ 8.70 (d, J=4.8 Hz, 2H), 7.60 (s, 1H), 7.37(d, J=8.0 Hz, 1H), 7.26-7.19 (m, 1H), 7.13-7.04 (m, 2H), 7.03-6.97 (m,1H), 6.86 (s, 1H), 6.69 (dd, J=9.8, 7.6 Hz, 1H), 6.46 (s, 1H), 6.07 (d,J=6.7 Hz, 1H), 4.87 (dd, J=9.1, 3.1 Hz, 1H), 4.36 (s, 1H), 3.11-2.83 (m,3H), 2.64-2.34 (m, 3H), 2.21 (m, 1H), 2.10-1.97 (m, 1H). MS: 576.1(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 71

¹H NMR (400 MHz, CDCl₃): δ 8.73 (d, J=7.1 Hz, 1H), 8.60-8.46 (m, 1H),7.56 (d, J=7.7 Hz, 1H), 7.38-7.32 (m, 1H), 7.31-7.27 (m, 1H), 7.26-7.18(m, 1H), 7.14-7.00 (m, 1H), 6.96 (m, 1H), 6.85 (s, 1H), 6.69 (m, 1H),6.40 (s, 1H), 6.02 (d, J=6.6 Hz, 1H), 4.98-4.74 (m, 1H), 4.39-4.10 (m,1H), 3.11-2.67 (m, 3H), 2.64-1.95 (m, 5H). MS: 600.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 86

¹H NMR (400 MHz, CDCl₃): δ 8.98 (d, J=4.8 Hz, 1H), 7.56 (s, 1H), 7.40(m, 2H), 7.23 (t, J=7.0 Hz, 1H), 7.08 (t, J=7.6 Hz, 1H), 7.01-6.84 (m,2H), 6.71 (t, J=8.6 Hz, 1H), 6.51 (s, 1H), 6.00 (d, J=6.7 Hz, 1H), 4.85(dd, J=9.3, 2.7 Hz, 1H), 4.36 (s, 1H), 3.15-2.80 (m, 3H), 2.67-2.26 (m,4H), 2.08 (dt, J=9.7, 8.1 Hz, 1H). MS: 601 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-5-oxo-N-(3-sulfamoylphenyl)pyrrolidine-2-carboxamide(Single Enantiomer)—Compound 53

¹H NMR (400 MHz, CDCl₃): δ 8.74 (s, 1H), 8.50 (s, 1H), 7.73 (d, J=7.5Hz, 1H), 7.33 (d, J=9.3 Hz, 1H), 7.25-6.80 (m, 6H), 6.40 (s, 1H), 5.61(d, J=6.9 Hz, 1H), 4.91 (d, J=8.0 Hz, 1H), 3.97 (s, 1H), 2.99-2.79 (m,1H), 2.55 (dd, J=13.7, 9.9 Hz, 1H), 2.25 (t, J=11.3 Hz, 1H), 2.03-1.74(m, 5H), 1.56-1.36 (m, 2H). MS: 610.2 (M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 81

¹H NMR (400 MHz, CDCl₃): δ 8.75 (s, 1H), 8.51 (d, J=5.0 Hz, 1H), 7.62(d, J=9.0 Hz, 1H), 7.37 (d, J=7.9 Hz, 1H), 7.27 (d, J=5.1 Hz, 1H), 7.22(t, J=7.7 Hz, 1H), 7.06 (t, J=7.5 Hz, 1H), 6.99 (d, J=6.9 Hz, 1H), 6.88(d, J=7.4 Hz, 1H), 6.69 (t, J=8.6 Hz, 1H), 6.41 (s, 1H), 5.69 (d, J=7.8Hz, 1H), 4.95 (dd, J=9.3, 3.2 Hz, 1H), 3.98 (m, 1H), 2.95-2.84 (m, 1H),2.65-2.55 (m, 1H), 2.30-2.20 (m, 1H), 2.05-2.12 (m, 1H), 2.03 (s, 2H),1.94-1.78 (m, 2H), 1.68-1.35 (m, 3H), 0.85-0.95 (m, 1H). MS: 628.2(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 87

¹H NMR (400 MHz, CDCl₃): δ 8.97 (d, J=4.8 Hz, 1H), 7.60 (d, J=8.7 Hz,1H), 7.46-7.34 (m, 2H), 7.22 (t, J=7.8 Hz, 1H), 7.06 (t, J=7.6 Hz, 1H),7.00-6.87 (m, 2H), 6.70 (t, J=8.6 Hz, 1H), 6.48 (s, 1H), 5.64 (d, J=7.7Hz, 1H), 4.86 (dd, J=9.3, 2.7 Hz, 1H), 3.98 (d, J=7.7 Hz, 1H), 2.96-2.86(m, 1H), 2.63-2.55 (m, 1H), 2.37-2.29 (m, 1H), 2.15-1.99 (m, 5H),1.96-1.77 (m, 2H), 1.61-1.34 (m, 2H). MS: 629.2 (M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-1-(2,4-dichlorophenyl)-2-((3,3-difluorocyclobutyl)-amino)-2-oxoethyl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 196

¹H NMR (400 MHz, CDCl₃): δ 8.77 (s, 1H), 8.49 (d, J=5.1 Hz, 1H), 7.56(s, 1H), 7.40 (d, J=2.1 Hz, 1H), 7.30 (s, 1H), 7.08 (dd, J=8.4, 2.1 Hz,1H), 6.97 (d, J=8.4 Hz, 1H), 6.90 (s, 1H), 6.79-6.72 (m, 1H), 6.35 (s,1H), 5.99 (d, J=6.6 Hz, 1H), 4.93 (dd, J=9.3, 3.1 Hz, 1H), 4.33 (s, 1H),3.12-2.95 (m, 2H), 2.95-2.83 (m, 1H), 2.66-2.32 (m, 3H), 2.24-2.18 (m,1H), 2.12-1.99 (m, 1H). MS: 634.1 (M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-1-(2,5-dichlorophenyl)-2-((3,3-difluorocyclobutyl)-amino)-2-oxoethyl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 201

¹H NMR (400 MHz, CDCl₃): δ 8.76 (s, 1H), 8.49 (dd, J=5.0, 0.6 Hz, 1H),7.58 (s, 1H), 7.30 (t, J=5.2 Hz, 2H), 7.22 (dd, J=8.6, 2.5 Hz, 1H), 7.02(d, J=2.4 Hz, 1H), 6.88 (s, 1H), 6.76 (tt, J=8.6, 2.3 Hz, 1H), 6.34 (s,1H), 6.14 (d, J=6.8 Hz, 1H), 4.94 (dd, J=9.3, 3.2 Hz, 1H), 4.43-4.28 (m,1H), 3.09-3.02 (m, 2H), 2.93-2.84 (m, 1H), 2.65-2.32 (m, 3H), 2.27-2.16(m, 1H), 2.14-2.00 (m, 1H). MS: 634.1 (M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-1-(2,6-dichlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 63

¹H NMR (400 MHz, CDCl₃): δ 8.77 (s, 1H), 8.45 (t, J=5.6 Hz, 1H), 7.88(t, J=10.0 Hz, 1H), 7.40-7.32 (m, 1H), 7.26-7.21 (m, 2H), 7.10-7.05 (m,2H), 6.92 (d, J=2.4 Hz, 1H), 6.62 (d, J=8.6 Hz, 1H), 5.53 (d, J=5.3 Hz,1H), 4.84-4.75 (m, 1H), 4.40 (s, 1H), 3.06-2.92 (m, 3H), 2.65-2.42 (m,4H), 2.18-2.02 (m, 1H). MS: 616.1 (M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-1-(2,6-dichlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 199

¹H NMR (400 MHz, CDCl₃): δ 8.78 (s, 1H), 8.44 (d, J=5.0 Hz, 1H),7.80-7.22 (m, 5H), 6.91 (s, 1H), 6.81 (tt, J=8.7, 2.3 Hz, 1H), 6.45 (d,J=8.5 Hz, 1H), 5.56 (d, J=6.8 Hz, 1H), 4.83 (dd, J=9.4, 2.7 Hz, 1H),4.40 (d, J=8.0 Hz, 1H), 3.23-2.92 (m, 3H), 2.69-2.39 (m, 4H), 2.23-2.02(m, 1H). MS: 634.2 (M+1)⁺.

(2S)-1-(4-Cyanopyridin-2-yl)-N-(1-(2,3-dichlorophenyl)-2-(3,3-difluoro-cyclobutylamino)-2-oxoethyl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 195

¹H NMR (400 MHz, CDCl₃): δ 8.72 (s, 1H), 8.57 (s, 1H), 7.44 (d, J=7.9,1H), 7.32-7.29 (m, 1H), 7.17-6.68 (m, 4H), 6.53-6.41 (m, 1H), 6.32-6.12(m, 1H), 4.90-4.65 (m, 1H), 4.41-4.05 (m, 1H), 3.13-2.01 (m, 8H). MS:634.1 (M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-2-(3,3-difluorocyclobutylamino)-1-(2-fluorophenyl)-2-oxoethyl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 208

¹H NMR (400 MHz, CDCl₃): δ 8.63 (s, 1H), 8.40 (d, J=4.9 Hz, 1H), 7.43(s, 1H), 7.20 (s, 1H), 7.16 (d, J=5.0 Hz, 1H), 6.90 (t, J=8.2 Hz, 3H),6.62 (t, J=8.7 Hz, 2H), 6.20 (s, 1H), 6.14 (d, J=6.4 Hz, 1H), 4.81 (dd,J=9.1, 2.9 Hz, 1H), 4.25 (s, 1H), 2.92 (s, 2H), 2.85-2.70 (m, 1H),2.56-2.22 (m, 3H), 2.15 (m, 1H), 2.04-1.90 (m, 1H). MS: 584.2 (M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-2-(3,3-difluorocyclobutylamino)-2-oxo-1-phenylethyl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 210

¹H NMR (400 MHz, CDCl₃): δ 8.77 (s, 1H), 8.41 (d, J=5.1 Hz, 1H), 7.49(s, 1H), 7.27 (dd, J=8.2, 5.0 Hz, 2H), 7.24 (d, J=5.4 Hz, 2H), 7.04 (d,J=6.7 Hz, 2H), 6.71 (t, J=8.8 Hz, 1H), 6.44 (s, 1H), 6.15 (s, 1H), 5.70(d, J=6.3 Hz, 1H), 4.86 (dd, J=9.3, 2.8 Hz, 1H), 4.29 (s, 1H), 2.99 (m,2H), 2.90 (m, 1H), 2.62-2.52 (m, 1H), 2.45 (m, 1H), 2.38-2.25 (m, 2H),2.07 (m, 1H). MS: 566.2 (M+1)⁺.

(S)—N—((S)-1-(3-Chloropyridin-2-yl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 198

¹H NMR (400 MHz, CDCl₃): δ 8.75 (s, 1H), 8.49 (d, J=5.0 Hz, 1H), 8.31(d, J=3.4 Hz, 1H), 7.65-7.56 (m, 2H), 7.27 (m, 1H), 7.19-7.15 (m, 1H),6.98 (m, 1H), 6.76-6.56 (m, 2H), 6.11 (d, J=6.8 Hz, 1H), 5.04-5.01 (m,1H), 4.38 (m, 1H), 3.05-2.98 (m, 2H), 2.92-2.83 (m, 1H), 2.60-2.52 (m,1H), 2.51-2.37 (m, 2H), 2.37-2.27 (m, 1H), 2.07-2.02 (m, 1H). MS: 601.1(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-5-oxo-N-(3-sulfamoylphenyl)pyrrolidine-2-carboxamide(Single Enantiomer)—Compound 84

¹H NMR (400 MHz, CDCl₃): δ 8.73 (d, J=10.0 Hz, 1H), 8.57-8.45 (d, J=8.0Hz, 1H), 8.12 (d, J=7.7 Hz, 1H), 7.83-7.76 (m, 2H), 7.61-7.56 (m, 1H),7.48-7.32 (m, 1H), 7.19 (t, J=7.1 Hz, 1H), 7.05-6.87 (m, 2H), 6.82-6.81(m, 1H), 6.55-6.43 (m, 1H), 6.27 (d, J=6.7 Hz, 1H), 5.24 (s, 1H), 4.84(d, J=7.2 Hz, 1H), 4.69 (s, 1H), 4.33 (s, 1H), 2.98-2.87 (m, 3H),2.63-2.24 (m, 4H), 2.09-2.00 (m, 1H). MS: 643.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-cyano-phenyl)-1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 128

¹H NMR (400 MHz, CDCl₃): δ 8.76 (s, 1H), 8.51 (s, 1H), 8.23 (m, 1H),7.58-7.27 (m, 4H), 6.93 (m, 3H), 6.43 (s, 1H), 5.85 (s, 1H), 4.78 (s,1H), 4.34 (s, 1H), 3.10-2.82 (m, 3H), 2.37-2.52 (m, 3H), 2.21-2.23 (m,1H), 1.89-1.99 (m, 1H). MS: 589.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-cyanophenyl)-1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide (SingleEnantiomer)—Compound 166

¹H NMR (400 MHz, CDCl₃): δ 8.77 (s, 1H), 8.49 (d, J=13.9 Hz, 1H),8.22-8.32 (m, 1H), 7.61-7.27 (m, 4H), 7.17-7.19 (m, 2H), 6.90-7.00 (m,2H), 6.42 (s, 1H), 5.50 (s, 1H), 4.80 (d, J=9.5 Hz, 1H), 3.97 (s, 1H),2.99-2.80 (m, 1H), 2.56-2.58 (m, 1H), 2.21-2.24 (m, 1H), 1.70-2.10 (m,6H), 1.41-1.44 (m, 2H). MS: 617.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-cyanophenyl)-1-(4-cyanopyrimidin-2-yl)-5-oxopyrrolidine-2-carboxamide (SingleEnantiomer)—Compound 167

¹H NMR (400 MHz, CDCl₃): δ 8.91-9.00 (m, 1H), 8.33-8.17 (m, 1H),7.62-7.32 (m, 5H), 7.20 (t, J=7.0 Hz, 1H), 7.02-7.06 (m, 1H), 6.95-6.83(m, 1H), 6.55 (s, 1H), 6.05-5.88 (m, 1H), 4.72 (d, J=9.3 Hz, 1H), 4.37(s, 1H), 2.91-3.05 (m, 3H), 2.70-2.25 (m, 4H), 2.13-1.92 (m, 1H). MS:590.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-cyanophenyl)-1-(4-cyanopyrimidin-2-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 178

¹H NMR (400 MHz, CDCl₃): δ 8.99 (s, 1H), 8.32 (s, 1H), 7.57 (m, 1H),7.54-7.28 (m, 2H), 7.19 (t, J=7.2 Hz, 3H), 7.04 (t, J=6.8 Hz, 1H), 6.93(d, J=7.7 Hz, 1H), 6.53 (s, 1H), 5.64-5.44 (m, 1H), 4.74 (d, J=9.3 Hz,1H), 3.99 (s, 1H), 2.94 (dd, J=17.8, 9.4 Hz, 1H), 2.62 (m, 1H),2.41-2.24 (m, 1H), 2.10-1.82 (m, 7H). MS: 618.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-cyano-5-fluorophenyl)-1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 177

¹H NMR (400 MHz, CDCl₃): δ 8.74 (s, 1H), 8.50 (s, 1H), 8.13-8.08 (m,1H), 7.44-7.27 (m, 2H), 7.23 (dd, J=12.6, 6.3 Hz, 2H), 7.07 (t, J=7.3Hz, 1H), 6.93 (t, J=6.4 Hz, 1H), 6.43 (d, J=6.1 Hz, 1H), 6.14 (dd,J=13.9, 6.7 Hz, 1H), 4.81 (dd, J=9.0, 2.3 Hz, 1H), 4.42-4.28 (m, 1H),3.12-2.94 (m, 2H), 2.94-2.80 (m, 1H), 2.67-2.29 (m, 3H), 2.23-1.92 (m,2H). MS: 607.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-N-(3-cyano-5-fluorophenyl)-1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 184

¹H NMR (400 MHz, CDCl₃): δ 8.77 (s, 1H), 8.50 (s, 1H), 8.25-8.03 (m,1H), 7.52-7.28 (m, 2H), 7.22 (t, J=7.7 Hz, 2H), 7.01 (dt, J=14.1, 10.1Hz, 2H), 6.42 (d, J=6.9 Hz, 1H), 5.58 (t, J=9.9 Hz, 1H), 4.83 (dd,J=9.1, 2.3 Hz, 1H), 4.05-3.86 (m, 1H), 3.04-2.81 (m, 1H), 2.59 (m, 1H),2.36-1.70 (m, 7H), 1.58-1.31 (m, 3H). MS: 636.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-cyano-5-fluorophenyl)-1-(4-cyanopyrimidin-2-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 185

¹H NMR (400 MHz, CDCl₃): δ 8.97 (d, J=4.4 Hz, 1H), 8.12 (m, 1H),7.50-7.32 (m, 3H), 7.23 (d, J=6.7 Hz, 2H), 7.06 (m, 1H), 6.95 (s, 1H),6.50 (d, J=8.6 Hz, 1H), 5.60 (d, J=7.5 Hz, 1H), 4.74 (d, J=8.8 Hz, 1H),3.98 (s, 1H), 2.90 (m, 1H), 2.72-2.49 (m, 1H), 2.28 (s, 1H), 2.17-1.67(m, 7H), 1.43 (m, 2H). MS: 637.2 (M+1)⁺.

(S)—N-(3-Cyano-5-fluorophenyl)-1-(4-cyanopyridin-2-yl)-N—((S)-2-(3,3-difluorocyclobutyl-amino)-2-oxo-1-phenylethyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 211

¹H NMR (400 MHz, CDCl₃): δ 8.71 (d, J=10.1 Hz, 1H), 8.38 (s, 1H), 8.02(m, 1H), 7.23 (m, 5H), 6.97 (d, J=7.3 Hz, 3H), 6.20 (s, 1H), 5.97 (s,1H), 4.70 (dd, J=9.2, 2.4 Hz, 1H), 4.27 (s, 1H), 2.93 (m, 2H), 2.85 (t,J=8.9 Hz, 1H), 2.59-2.48 (m, 1H), 2.49-2.29 (m, 2H), 2.29-2.20 (m, 1H),2.08-1.99 (m, 1H). MS: 573.2 (M+1)⁺.

(S)—N-(3-Cyano-5-fluorophenyl)-1-(4-cyanopyridin-2-yl)-N—((S)-2-((3,3-difluorocyclobutyl)amino)-1-(2-fluorophenyl)-2-oxoethyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 207

¹H NMR (400 MHz, DMSO-d₆): δ 8.78 (s, 1H), 8.62 (d, J=5.1 Hz, 1H), 8.48(s, 1H), 8.04-7.83 (m, 1H), 7.78 (s, 1H), 7.57 (s, 1H), 7.23 (m, 2H),7.14 (d, J=9.9 Hz, 1H), 6.95 (t, J=7.5 Hz, 1H), 6.84 (s, 1H), 6.20 (s,1H), 4.72 (s, 1H), 4.04 (s, 1H), 4.00-3.82 (m, 1H), 3.09-2.67 (m, 2H),2.33 (m, 1H), 1.91 (s, 2H), 1.83 (s, 1H), 1.27-1.05 (m, 1H). MS: 591.2(M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 91

¹H NMR (400 MHz, CDCl₃): δ 9.10-8.03 (m, 4H), 7.47-7.39 (m, 2H),7.27-6.84 (m, 3H), 6.51-6.01 (m, 2H), 4.84-4.70 (m, 1H), 4.36-4.20 (m,1H), 3.25-1.86 (m, 8H). MS: 583.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 176

¹H NMR (400 MHz, CDCl₃): δ 8.95-8.70 (m, 1H), 8.49 (d, J=4.7 Hz, 1H),8.36-8.11 (m, 1H), 8.12 (d, J=8.6 Hz, 1H), 7.33 (d, J=8.0 Hz, 1H), 7.21(t, J=7.8 Hz, 1H), 7.04 (t, J=7.6 Hz, 1H), 6.48-6.41 (m, 1H), 6.30-6.21(m, 1H), 4.84-6.79 (m, 1H), 4.38-4.30 (m, 1H), 3.11-2.74 (m, 3H),2.65-1.91 (m, 5H). MS: 583.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 193

¹H NMR (400 MHz, CDCl₃): δ 8.77 (s, 1H), 8.49 (d, J=5.2 Hz, 1H),8.40-8.27 (m, 1H), 8.21-8.04 (m, 1H), 7.41-7.36 (m, 1H), 7.26-7.23 (m,1H), 7.20 (t, J=6.9 Hz, 1H), 7.04 (t, J=7.2 Hz, 1H), 6.93 (m, 1H),6.52-6.34 (m, 1H), 5.49 (s, 1H), 4.84 (d, J=7.4 Hz, 1H), 4.01-3.94 (m,1H), 2.99-2.91 (m, 1H), 2.62-2.54 (m, 1H), 2.22-1.71 (m, 7H), 1.31 (s,3H). MS: 611.2 (M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-2-((3,3-difluorocyclobutyl)amino)-2-oxo-1-phenylethyl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 147

¹H NMR (400 MHz, CDCl₃): δ 8.86 (m, 1H), 8.39 (m, 2H), 8.03 (m, 1H),7.28 (d, J=5.9 Hz, 4H), 6.98 (m, 2H), 6.29 (s, 1H), 5.85 (s, 1H), 4.85(m, 1H), 4.33 (s, 1H), 3.26-2.82 (m, 3H), 2.69-1.88 (m, 5H). MS: 549.2(M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-2-((3,3-difluorocyclobutyl)amino)-1-(2-fluorophenyl)-2-oxoethyl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 148

¹H NMR (400 MHz, CDCl₃): δ 8.99-8.60 (m, 1H), 8.55-7.97 (m, 3H),7.35-7.19 (m, 3H), 7.07-6.89 (m, 3H), 6.36 (m, 1H), 6.12 (s, 1H), 4.80(s, 1H), 4.35 (s, 1H), 3.22-2.79 (m, 3H), 2.64-1.85 (m, 5H). MS: 567.2(M+1)⁺.

(S)-1-(4-Cyanopyridin-2-yl)-N—((S)-2-((3,3-difluorocyclobutyl)amino)-2-oxo-1-phenylethyl)-N-(5-isocyanopyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 212

¹H NMR (400 MHz, CDCl₃): δ 9.34 (s, 1H), 8.87-8.56 (m, 4H), 8.41 (s,2H), 8.27 (s, 1H), 7.54 (s, 7H), 7.01 (d, J=6.9 Hz, 3H), 6.35 (s, 2H),5.73 (s, 2H), 4.66 (s, 2H), 4.35 (s, 2H), 2.99 (m, 5H), 2.73-2.20 (m,7H), 2.07 (s, 2H). MS: 556.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(3-cyano-phenyl)-N-(1H-indazol-7-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 186

¹H NMR (400 MHz, CDCl₃): δ 8.72-8.71 (m, 1H), 8.66 (s, 1H), 8.08 (s,1H), 7.69 (s, 1H), 7.67 (s, 1H), 7.50-7.49 (m, 1H), 7.36-7.34 (m, 1H),7.11-7.07 (m, 1H), 7.00-6.96 (m, 1H), 6.83-6.76 (m, 2H), 6.48 (s, 1H),5.07-5.07 (m, 1H), 4.38-4.33 (m, 1H), 3.05-2.91 (m, 2H), 2.80-2.71 (m,1H), 2.65-2.60 (m, 1H), 2.53-2.46 (m, 2H), 2.03-1.99 (m, 1H), 1.75-1.67(m, 1H). MS: 603.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-3-(3-cyanophenyl)-N-(1H-indazol-7-yl)-2-oxooxazolidine-4-carboxamide (SingleEnantiomer)—Compound 142

¹H NMR (400 MHz, CDCl₃): δ 13.03 (s, 1H), 8.73 (s, 1H), 8.55-8.54 (m,1H), 8.02 (s, 1H), 8.58-8.56 (m, 1H), 8.50-8.48 (m, 1H), 7.27-7.24 (m,2H), 7.03-6.99 (m, 1H), 6.91-6.87 (m, 1H), 6.80-6.78 (m, 1H), 6.72-6.68(m, 1H), 6.33 (s, 2H), 5.70-5.69 (m, 1H), 4.99-4.97 (m, 1H), 4.05-4.03(m, 1H), 2.78-2.95 (m, 1H), 2.47-2.40 (m, 1H), 2.08-4.99 (m, 6H),1.90-1.82 (m, 2H), 1.67-1.63 (m, 1H), 1.58-1.62 (m, 1H). MS: 633.2(M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(1H-indazol-4-yl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 152

¹H NMR (400 MHz, DMSO-d₆): δ 13.05 (m, 1H), 8.70 (m, 2H), 8.54 (d, J=6.7Hz, 1H), 8.21 (s, 1H), 7.80 (d, J=6.9 Hz, 1H), 7.63 (d, J=5.0 Hz, 1H),7.36 (m, 2H), 7.24 (d, J=8.0 Hz, 1H), 7.18-6.97 (m, 1H), 6.92-6.79 (m,1H), 6.77-6.70 (m, 1H), 6.35 (d, 1H), 4.66 (m, 1H), 4.20-4.01 (m, 1H),3.05-2.78 (m, 2H), 2.68-2.52 (m, 2H), 2.49-2.26 (m, 2H), 2.22-1.53 (m,2H). MS: 604.2 (M+1)⁺.

(S)—N-(3-(1H-Pyrazol-4-yl)phenyl)-N—((S)-1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 200

¹H NMR (400 MHz, MeOD): δ 8.73-8.54 (m, 2H), 8.14-7.91 (m, 1H), 7.71 (d,J=7.6 Hz, 1H), 7.56-7.28 (m, 4H), 7.25-6.92 (m, 4H), 6.70 (d, J=7.6 Hz,1H), 6.54-6.39 (m, 1H), 5.03 (dd, J=9.4, 2.9 Hz, 1H), 4.31-4.05 (m, 1H),3.00-2.73 (m, 3H), 2.64-2.00 (m, 5H). MS: 630.2 (M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-5-oxo-N-(3-(trifluoromethoxy)phenyl)pyrrolidine-2-carboxamide(Racemic)—Compound 180

¹H NMR (400 MHz, CDCl₃): δ 8.96 (t, J=5.5 Hz, 1H), 7.88 (s, 1H),7.44-7.32 (m, 2H), 7.21 (m, 2H), 7.10 (t, J=7.3 Hz, 1H), 7.04-6.95 (m,1H), 6.91 (m, 1H), 6.52 (m, 1H), 6.18 (m, 1H), 4.89-4.67 (m, 1H), 4.31(m, 1H), 3.22-2.75 (m, 3H), 2.70-1.92 (m, 5H). MS: 649.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(difluoromethoxy)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 181

¹H NMR (400 MHz, CDCl₃): δ 8.74 (s, 1H), 8.44 (m, 1H), 7.76 (d, J=9.0Hz, 1H), 7.33 (m, 2H), 7.21-6.83 (m, 6H), 6.44 (t, J=8.8 Hz, 1H),6.28-6.13 (m, 1H), 4.91 (m, 1H), 4.34 (s, 1H), 3.10-2.66 (m, 3H),2.65-1.84 (m, 5H). MS: 630.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-(difluoromethoxy)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 194

¹H NMR (400 MHz, CDCl₃): δ 9.04-8.59 (m, 1H), 7.74 (s, 1H), 7.43-7.26(m, 4H), 6.96 (m, 3H), 6.36 (m, 2H), 4.81 (t, J=9.3 Hz, 1H), 4.55 (m,1H), 4.33 (s, 1H), 4.06-3.89 (m, 1H), 3.15-2.69 (m, 2H), 2.69-1.86 (m,5H). MS: 631.1 (M+1)⁺.

(S)—N—((S)-1-(2C)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-methoxyphenyl)-5-oxopyrrolidine-2-carboxamide (SingleEnantiomer)—Compound 129

¹H NMR (400 MHz, CDCl₃): δ 8.75 (s, 1H), 8.51 (d, J=5.0 Hz, 1H), 7.47(m, 1H), 7.38-7.08 (m, 3H), 6.99 (d, J=6.7 Hz, 3H), 6.89-6.66 (m, 2H),6.41 (s, 1H), 6.09 (d, J=6.6 Hz, 1H), 4.97 (dd, J=9.3, 3.2 Hz, 1H), 4.34(s, 1H), 3.72 (m, 3H), 3.01 (dd, J=7.5, 4.0 Hz, 3H), 2.65-2.23 (m, 4H),2.04 (d, J=9.0 Hz, 1H). MS: 594.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutyl-amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-methoxyphenyl)-5-oxopyrrolidine-2-carboxamid(Single Enantiomer)—Compound 164

¹H NMR (400 MHz, CDCl₃): δ 8.92 (s, 1H), 7.48-7.39 (m, 1H), 7.33-7.26(m, 2H), 7.22-7.08 (m, 2H), 7.04-6.82 (m, 3H), 6.73 (s, 2H), 6.48 (d,J=9.5 Hz, 1H), 6.18 (m, 1H), 4.88-4.85 (m, 1H), 4.32 (s, 1H), 3.78 (s,1H), 3.62 (s, 2H), 3.01-2.81 (m, 3H), 2.58-2.49 (m, 2H), 2.42-2.30 (m,2H), 2.09-1.98 (m, 1H). MS: 595 (M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-cyclopropoxyphenyl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 192

¹H NMR (400 MHz, CDCl₃): δ 9.06-8.88 (m, 1H), 7.61-7.30 (m, 4H),7.27-7.22 (m, 1H), 7.18 (t, J=7.4 Hz, 2H), 7.08-6.92 (m, 1H), 6.87 (dd,J=8.7, 2.1 Hz, 1H), 6.78 (t, J=9.5 Hz, 1H), 6.50 (s, 1H), 6.04 (m, 3H),5.57-5.14 (m, 2H), 4.88 (m, 1H), 4.77-4.10 (m, 3H), 3.15-2.75 (m, 3H),2.68-2.47 (m, 2H), 2.45-2.21 (m, 3H), 2.20-1.90 (m, 1H). MS: 621.1(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(hydroxymethyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 131

¹H NMR (400 MHz, CDCl₃): δ 8.73 (s, 1H), 8.53 (s, 1H), 7.94-7.70 (m,1H), 7.31 (s, 1H), 7.26 (dd, J=5.1, 1.3 Hz, 1H), 7.22-7.10 (m, 4H),7.02-6.87 (m, 2H), 6.44 (d, J=10.5 Hz, 1H), 6.12 (d, J=6.4 Hz, 1H), 4.91(dd, J=9.3, 3.2 Hz, 1H), 4.69 (s, 1H), 4.48 (s, 1H), 4.42-4.26 (m, 1H),3.07-2.85 (m, 3H), 2.65-2.17 (m, 4H), 2.01 (s, 2H). MS: 594.2 (M+1)⁺.

(S)—N—((S)-1-(2-Xhlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(1-hydroxycyclopropyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 140

¹H NMR (400 MHz, CDCl₃): δ ¹H NMR (400 MHz, CDCl₃): δ 8.73 (s, 1H),8.52-8.44 (m, 1H), 7.64-7.30 (m, 3H), 7.22-6.90 (m, 5H), 6.42-6.38 (m,1H), 6.03 (m, 1H), 4.87 (m, 1H), 4.30 (m, 1H), 3.05-2.82 (m, 3H),2.60-1.88 (m, 5H), 1.21 (d, J=3.2 Hz, 4H). MS: 620.2 (M+1)⁺.

(S)—N—((S)-1-(2C)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(2-hydroxypropan-2-yl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 179

¹H NMR (400 MHz, CDCl₃): δ 8.69 (s, 1H), 8.54 (d, J=5.0 Hz, 1H),7.93-7.70 (m, 1H), 7.40-7.19 (m, 4H), 7.11 (m, 2H), 7.01-6.72 (m, 2H),6.45 (m, 2H), 5.05-4.76 (m, 1H), 4.33 (s, 1H), 3.13-2.58 (m, 3H), 2.42(m, 4H), 2.09-1.83 (m, 1H), 1.33 (s, 6H). MS: 622.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluoro-5-(2-hydroxypropan-2-yl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 150

¹H NMR (400 MHz, CDCl₃): δ 8.66 (s, 1H), 8.49 (d, J=4.8 Hz, 1H),7.73-7.48 (m, 1H), 7.26-6.83 (m, 7H), 6.53-6.42 (m, 2H), 4.91 (d, J=6.4Hz, 1H), 4.32 (s, 1H), 3.02-2.72 (m, 3H), 2.58-1.85 (m, 6H), 1.63 (s,2H), 1.51 (d, J=7.0 Hz, 2H), 1.29 (d, J=8.6 Hz, 4H). MS: 640.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluoro-5-(2-hydroxypropan-2-yl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 155

¹H NMR (400 MHz, CDCl₃): δ 8.80 (s, 1H), 8.43 (s, 1H), 7.51 (d, 1H),7.24 (m, 4H), 7.06 (s, 3H), 6.64 (m, 1H), 6.15 (m, 1H), 5.73 (s, 1H),4.86 (s, 1H), 4.32 (s, 1H), 3.01 (m, 3H), 2.68-2.27 (m, 4H), 2.12 (s,1H), 1.44 (s, 1H), 1.29 (d, J=9.0 Hz, 6H). MS: 639.2 (M+1)⁺

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(2-hydroxyethyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 160

¹H NMR (400 MHz, CDCl₃): δ 8.76 (s, 1H), 8.52 (d, J=5.0 Hz, 1H), 7.74(s, 1H), 7.32-7.36 (m, 1H), 7.27-7.11 (m, 2H), 7.09-6.87 (m, 4H),6.39-6.45 (m, 1H), 6.05 (d, J=6.9 Hz, 1H), 4.33 (s, 1H), 3.82 (s, 1H),3.59 (s, 1H), 3.12-2.79 (m, 4H), 2.74-2.16 (m, 5H), 1.99-2.07 (m, 1H).MS: 608.2 (M+1)⁺.

(S)—N—((S)-1-(2-Clorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(2-hydroxyethoxy)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 130

¹H NMR (400 MHz, CDCl₃): δ 8.72 (s, 1H), 8.48 (d, J=5.0 Hz, 1H),7.54-7.28 (m, 2H), 7.18-7.21 (m, 2H), 7.01-6.94 (m, 2H), 6.75-6.77 (m,2H), 6.39 (s, 1H), 5.99 (s, 1H), 4.94 (dd, J=9.3, 3.4 Hz, 1H), 4.31 (s,1H), 3.79-4.06 (m, 4H), 3.07-2.80 (m, 3H), 2.58-2.21 (m, 4H), 1.87-2.00(m, 2H). MS: 624.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluoro-54(S)-methylsulfinyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 190

¹H NMR (400 MHz, CDCl₃): δ 8.75 (s, 1H), 8.54 (m, 1H), 8.02-7.78 (m,1H), 7.33 (s, 3H), 7.21 (m 1H), 7.06 (t, J=7.4 Hz, 1H), 6.96 (m, 1H),6.45 (m, 1H), 6.27 (m, 1H), 4.86 (m, 1H), 4.35 (m, 1H), 3.16-2.82 (m,3H), 2.71 (s, 1H), 2.65-2.47 (m, 2H), 2.41 (m, 3H), 2.22 (m, 1H), 2.09(m, 1H). MS: 644.1 (M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 96

¹H NMR (400 MHz, CDCl3): δ 8.84-8.11 (m, 3H), 7.93-7.35 (m, 4H),7.25-6.75 (m, 2H), 6.64-5.94 (m, 2H), 4.89-4.69 (m, 1H), 4.28 (d, J=5.7Hz, 1H), 3.13-2.74 (m, 6H), 2.68-2.48 (m, 2H), 2.46-2.15 (m, 3H), 2.04(s, 1H). MS: 642.1 (M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 102

¹H NMR (400 MHz, CDCl₃): δ 8.93 (t, J=5.3 Hz, 1H), 8.50-8.15 (m, 1H),7.94-7.71 (m, 2H), 7.66-7.46 (m, 1H), 7.38 (t, J=6.4 Hz, 1H), 7.28 (t,J=3.6 Hz, 1H), 7.20-7.07 (m, 1H), 7.05-6.87 (m, 2H), 6.74 (m, 1H), 6.52(m, 1H), 4.72 (dd, J=9.2, 2.5 Hz, 1H), 4.34 (d, J=6.4 Hz, 1H), 3.00 (s,3H), 2.90-2.75 (m, 3H), 2.56-2.19 (m, 5H), 1.98 (m, 1H). MS: 643.1(M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 95

¹H NMR (400 MHz, CDCl₃): δ 8.87-8.13 (m, 3H), 8.02-7.37 (m, 4H),7.24-6.87 (m, 2H), 6.51-6.39 (m, 1H), 5.77-5.28 (m, 1H), 4.89-4.65 (m,1H), 3.94 (d, J=5.2 Hz, 1H), 3.16-2.73 (m, 4H), 2.68-2.53 (m, 1H),2.44-2.20 (m, 1H), 2.03 (m, 8H), 1.44 (m, 2H). MS: 670.2 (M+1)⁺.

(S)—N—((S)-1-(2C)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 103

¹H NMR (400 MHz, CDCl₃): δ 8.94 (dd, J=7.9, 4.8 Hz, 1H), 8.56-8.15 (m,1H), 7.97-7.62 (m, 2H), 7.56-7.29 (m, 3H), 7.13 (t, J=7.6 Hz, 1H),7.06-6.84 (m, 2H), 6.51 (d, J=4.2 Hz, 1H), 6.10 (dd, J=3.2, 7.4 Hz, 1H),4.74 (d, J=6.6 Hz, 1H), 3.98 (s, 1H), 3.01 (s, 1H), 2.93-2.72 (m, 3H),2.52 (d, J=9.6 Hz, 1H), 2.37-2.20 (m, 1H), 2.13-1.78 (m, 7H), 1.63-1.40(m, 2H). MS: 671 (M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluoro-5-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 110

¹H NMR (400 MHz, CDCl₃): δ 8.45-8.79 (m, 2H), 8.40-8.13 (s, 1H),8.09-7.67 (m, 1H), 7.63-7.30 (m, 2H), 7.23-6.87 (m, 3H), 6.55-6.30 (m,1H), 6.22-5.94 (m, 1H), 4.96-4.61 (m, 1H), 4.26 (m, 4H), 3.16-1.87 (m,7H), 1.27 (d, 1H). MS: 660.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-fluoro-5-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 109

¹H NMR (400 MHz, CDCl₃): δ 8.96 (d, J=4.6 Hz, 3H), 7.99 (d, J=8.5 Hz,2H), 7.75 (s, 2H), 7.52 (d, J=7.0 Hz, 3H), 7.37 (d, J=4.9 Hz, 5H), 7.19(t, J=7.7 Hz, 3H), 7.01 (dt, J=7.1 Hz, 6H), 6.40-6.60 (m, 3H), 6.06 (d,J=6.5 Hz, 3H), 4.76 (d, J=9.2 Hz, 1H), 4.35 (m, 4H), 3.14-1.87 (m, 8H).MS: 661.1 (M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluoro-5-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 105

¹H NMR (400 MHz, CDCl₃): δ 8.96 (t, J=4.6 Hz, 1H), 7.53-7.36 (m, 3H),7.23 (m, J=7.8, 1.5 Hz, 1H), 7.14-6.94 (m, 3H), 6.68 (m, J=8.6, 2.3 Hz,1H), 6.60 (d, J=3.1 Hz, 1H), 6.07 (d, J=6.7 Hz, 1H), 4.75 (q, J=4.0, 2.1Hz, 1H), 4.38 (d, J=6.7 Hz, 1H), 3.78-3.67 (m, 2H), 3.39 (m, 1H),3.26-2.92 (m, 3H), 2.67-2.36 (m, 2H). MS: 688.1 (M+1)⁺.

(2S)—N-(1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-fluoro-5-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 108

¹H NMR (400 MHz, CDCl₃): δ 8.97 (s, 1H), 8.20-8.60 (m, 1H), 8.09-7.68(m, 1H), 7.63-7.32 (m, 5H), 7.22-6.93 (m, 3H), 6.64-6.03 (m, 2H), 5.62(s, 1H), 4.60-4.85 (m, 1H), 3.21-1.70 (m, 12H), 1.50-1.14 (m, 2H). MS:689.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-fluoro-5-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 168

¹H NMR (400 MHz, CDCl₃): δ 9.0 (s, 1H), 8.05-8.02 (m, 1H), 7.80 (m, 1H),7.56-7.00 (m, 7H), 6.58 (m, 1H), 5.65 (m, 1H), 4.80 (m, 1H), 4.14 (m,1H), 3.00-0.88 (m, 15H). MS: 689.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(methylsulfonamido)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 159

¹H NMR (400 MHz, DMSO-d₆): δ 9.78 (s, 1H), 8.84-8.61 (m, 2H), 8.56 (s,1H), 7.66 (m, 2H), 7.49-7.15 (m, 3H), 7.15-6.79 (m, 4H), 6.25 (m, 1H),4.89-4.74 (m, 1H), 4.19-4.04 (m, 1H), 3.03-2.83 (m, 3H), 2.72-2.59 (m,3H), 2.54 (m, 2H), 2.44-2.28 (m, 1H), 1.99 (m, 2H). MS: 657.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(dimethylamino)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 161

¹H NMR (400 MHz, CDCl₃): δ 8.71 (d, J=9.9 Hz, 1H), 8.50-8.41 (m, 1H),7.29 (d, J=7.8 Hz, 1H), 7.22 (dd, J=5.0, 1.3 Hz, 1H), 7.18-7.05 (m, 2H),6.99-6.86 (m, 3H), 6.56-6.47 (m, 2H), 6.37 (d, J=6.6 Hz, 1H), 6.11 (s,1H), 5.01 (d, J=9.2 Hz, 1H), 4.34-4.28 (m, 1H), 3.07-2.70 (m, 8H),2.61-2.42 (m, 2H), 2.35-2.25 (m, 2H), 2.01-1.97 (m, 1H). MS: 607.2(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(2-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 187

¹H NMR (400 MHz, CDCl₃): δ 8.74 (m, 1H), 8.48 (m, 1H), 7.96-7.92 (m,1H), 7.40 (m, 1H), 7.28-6.72 (m, 7H), 6.59-5.79 (m, 2H), 4.86-4.78 (m,1H), 4.28 (s, 1H), 3.04-2.90 (m, 3H), 2.66-2.01 (m, 5H). MS: 582.1(M+1)⁺.

(S)—N—((S)-1-(2-Clorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(2,3-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 188

¹H NMR (400 MHz, CDCl₃): δ 8.73 (m, 1H), 8.47 (d, J=5.0 Hz, 1H),7.84-7.73 (m, 1H), 7.43 (d, J=8.1 Hz, 1H), 7.28-7.20 (m, 2H), 7.13 (dd,J=8.2, 4.4 Hz, 2H), 7.01-6.83 (m, 2H), 6.62 (s, 1H), 6.42-5.85 (m, 1H),4.85-4.77 (m, 1H), 4.20 (m, 1H), 3.13-2.78 (m, 3H), 2.68-2.28 (m, 4H),2.25-2.04 (m, 1H). MS: 600.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(2,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 197

¹H NMR (400 MHz, CDCl₃): δ 8.73 (m, 1H), 8.54-8.41 (m, 1H), 7.83-7.78(m, 1H), 7.44-7.39 (m, 1H), 7.28-7.21 (m, 2H), 7.13-6.88 (m, 3H),6.81-6.80 (m, 1H), 6.61-6.31 (m, 1H), 5.91 (d, J=6.5 Hz, 1H), 4.86-4.79(m, 1H), 4.29 (dd, J=8.2, 6.7 Hz, 1H), 3.51 (s, 1H), 3.12-2.85 (m, 3H),2.68-2.56 (m, 1H), 2.54-2.45 (m, 1H), 2.43-2.24 (m, 2H), 2.23-2.06 (m,1H). MS: 600.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(1H-indazol-5-yl)-5-oxopyrrolidine-2-carboxamide (SingleEnantiomer)—Compound 203

¹H NMR (400 MHz, CDCl₃): δ 8.78 (s, 1H), 8.56 (m, 1H), 8.39 (s, 1H),8.13-7.88 (m, 1H), 7.44-7.32 (m, 2H), 7.28-7.00 (m, 4H), 6.99-6.79 (m,2H), 6.48 (m, 1H), 5.75-5.48 (m, 1H), 5.06-4.75 (m, 1H), 4.00 (s, 1H),3.10-2.77 (m, 1H), 2.63-2.44 (m, 1H), 2.37-2.20 (m, 1H), 2.15-1.77 (m,7H), 1.42 (m, 2H). MS: 632.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(1H-indazol-6-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 205

¹H NMR (400 MHz, CDCl₃): δ 8.78 (s, 1H), 8.57 (t, J=5.0 Hz, 1H),8.23-7.76 (m, 2H), 7.54-7.30 (m, 2H), 7.16 (s, 1H), 7.04-6.86 (m, 3H),6.47 (d, J=11.7 Hz, 1H), 6.02 (d, J=6.1 Hz, 1H), 4.92 (m, 1H), 4.36 (s,1H), 2.97 (m, 3H), 2.65-2.20 (m, 4H), 1.99 (m, 1H). MS: 604.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(1H-indazol-6-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 136

¹H NMR (400 MHz, CDCl₃): δ 10.41-9.94 (m, 1H), 8.79 (s, 1H), 8.57 (t,J=5.1 Hz, 1H), 8.28-8.09 (m, 1H), 7.93 (m, 1H), 7.52 (s, 1H), 7.40 (d,J=7.9 Hz, 1H), 7.33 (d, J=7.5 Hz, 1H), 7.15-6.98 (m, 1H), 6.46 (d,J=12.7 Hz, 1H), 5.50 (d, J=7.9 Hz, 1H), 5.06-4.76 (m, 1H), 4.02 (s, 1H),2.92 (dd, 1H), 2.63-2.49 (m, 1H), 2.31 (s, 1H), 2.03 (m, 6H), 1.45 (s,2H). MS: 632.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(1H-indazol-5-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 175

¹H NMR (400 MHz, CDCl₃): δ 8.75 (s, 1H), 8.64-8.46 (m, 1H), 8.34 (s,1H), 8.09 (s, 1H), 7.94-7.92 (m, 1H), 7.42-7.32 (m, 2H), 7.24-7.02 (m,2H), 6.94-6.85 (m, 2H), 6.49-6.45 (m, 1H), 6.08-6.06 (m, 1H), 5.00-4.76(m, 1H), 4.35-4.31 (s, 1H), 3.00-2.85 (m, 3H), 2.64-2.11 (m, 4H),2.01-1.93 (m, 1H). MS: 604.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(1H-indol-5-yl)-5-oxopyrrolidine-2-carboxamide (SingleEnantiomer)—Compound 206

¹H NMR (400 MHz, CDCl₃): δ 8.74 (s, 1H), 8.55 (m, 1H), 8.12 (d, J=13.8Hz, 2H), 7.52-7.29 (m, 2H), 7.18-6.80 (m, 5H), 6.46 (m, 2H), 5.83 (s,1H), 5.83 (s, 1H), 5.08-4.81 (m, 1H), 4.33 (s, 1H), 2.92 (m, 3H),2.64-2.16 (m, 4H), 2.01 (m, 1H). MS: 603.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(1-methyl-1H-indol-5-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 209

¹H NMR (400 MHz, CDCl₃): δ 8.83-8.39 (m, 1H), 8.01 (m, 1H), 7.68-7.32(m, 1H), 7.28-6.72 (m, 8H), 6.55-6.38 (m, 1H), 5.90 (m, 1H), 5.00-4.73(m, 1H), 4.33 (s, 1H), 3.80-3.62 (m, 3H), 2.91 (m, 3H), 2.62-1.78 (m,5H). MS: 617.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-cyclopropylphenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 173

¹H NMR (400 MHz, CDCl₃): δ 8.76 (s, 1H), 8.59 (d, J=4.8 Hz, 1H),7.50-7.60 (m, 1H), 7.41 (d, J=7.8 Hz, 1H), 7.28-7.19 (m, 2H), 7.14-6.94(m, 2H), 6.62-6.79 (m, 1H), 6.26-6.07 (m, 2H), 4.86 (dd, J=9.3, 2.9 Hz,1H), 4.16-4.19 (m, 1H), 3.02-2.76 (m, 3H), 2.57-2.59 (m, 1H), 2.40-2.16(m, 3H), 2.02-2.12 (m, 1H), 1.28-1.29 (m, 2H), 0.90 (t, J=6.9 Hz, 2H).MS: 604.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-cyclopropylphenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 182

¹H NMR (400 MHz, CDCl₃): δ 8.94 (d, J=4.5 Hz, 1H), 7.57-7.49 (m, 1H),7.43-7.28 (m, 2H), 7.19-7.14 (m, 2H), 7.05-6.79 (m, 4H), 6.51-6.46 (m,1H), 6.00-5.97 (m, 1H), 4.82-4.80 (m, 1H), 4.32-4.33 (m, 1H), 3.09-2.81(m, 3H), 2.64-2.24 (m, 4H), 2.05-1.72 (m, 2H), 0.99-0.76 (m, 4H). MS:605.2 (M+1)⁺.

(S)—N-(3-(tert-Butyl)phenyl)-N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 165

¹H NMR (400 MHz, CDCl₃): δ 8.94 (d, J=4.8 Hz, 1H), 8.00-7.54 (m, 1H),7.41-7.32 (m, 2H), 7.24-7.15 (m, 2H), 7.14-7.02 (m, 2H), 6.97-6.81 (m,2H), 6.53 (s, 1H), 6.20 (dd, J=12.7, 6.8 Hz, 1H), 4.86 (m, 1H), 4.34 (s,1H), 3.15-2.80 (m, 3H), 2.63-2.27 (m, 4H), 2.13-1.92 (m, 1H), 1.29 (s,9H). MS: 621.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-cyclopropyl-5-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 204

¹H NMR (400 MHz, CDCl₃): δ 8.77 (s, 1H), 8.50 (s, 1H), 7.50-7.33 (m,2H), 7.24-7.17 (m, 1H), 7.01 (m, 2H), 6.68 (m, 2H), 6.39 (m, 1H), 6.00(s, 1H), 4.93 (s, 1H), 4.34 (s, 1H), 3.15-2.83 (m, 3H), 2.59-2.53 (m,2H), 2.40-2.37 (m, 2H), 2.07 (s, 1H), 1.27 (s, 1H), 1.05 (s, 1H), 0.91(d, J=6.7 Hz, 1H), 0.67 (s, 1H), 0.43 (m, 1H). MS: 622.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-cyclopropyl-5-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 202

¹H NMR (400 MHz, CDCl₃): δ 8.79 (s, 1H), 8.50 (s, 1H), 7.40 (m, 2H),7.15 (m, 1H), 7.01 (m, 3H), 6.84-6.56 (m, 2H), 6.38 (m, 1H), 5.50 (s,1H), 4.94 (s, 1H), 3.99 (s, 1H), 2.90 (m, 1H), 2.57 (m, 1H), 2.28 (s,1H), 2.05 (m, 5H), 1.92-1.77 (m, 2H), 1.30 (m, 2H), 0.91 (t, J=6.7 Hz,2H), 0.67 (s, 2H). MS: 650.2 (M+1)⁺.

((S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(N-methylsulfamoyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 157

¹H NMR (400 MHz, CD₃OD): δ 8.89-8.59 (m, 3H), 8.50-8.01 (m, 2H),7.69-7.31 (m, 5H), 7.17 (t, J=7.6 Hz, 2H), 7.03 (t, J=7.6 Hz, 2H), 6.95(t, J=7.9 Hz, 2H), 6.51 (s, 1H), 4.98 (s, 1H), 4.24 (s, 2H), 3.01-2.45(m, 7H), 2.35 (s, 3H), 2.10-2.05 (m, 1H). MS: 657.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-(N,N-dimethylsulfamoyl)phenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 156

¹H NMR (400 MHz, CDCl₃): δ 8.70 (s, 1H), 8.60 (d, J=4.9 Hz, 1H), 8.17(d, J=7.7 Hz, 1H), 7.86 (s, 1H), 7.63-7.55 (m, 1H), 7.49 (t, J=7.8 Hz,1H), 7.27 (s, 1H), 7.20-6.92 (m, 4H), 6.50 (d, J=6.9 Hz, 2H), 4.79 (d,J=7.0 Hz, 1H), 4.32 (s, 1H), 3.05-2.75 (m, 4H), 2.60-1.90 (m, 10H). MS:671.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(3-cyanopyridin-2-yl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 69

¹H NMR (400 MHz, CDCl₃): δ 8.14 (d, J=8.0 Hz, 1H), 7.93 (d, J=4.0 Hz,1H), 7.92 (m, 1H), 7.17-7.28 (m, 4H), 6.91-7.04 (m, 4H), 6.42 (s, 1H),6.31 (s, 1H), 4.87-4.91 (m, 1H), 4.35 (m, 1H), 2.97-3.02 (m, 2H),2.79-2.86 (m, 1H), 2.45-2.57 (m, 3H), 2.23-2.26 (m, 1H), 2.09-2.11 (m,1H). MS: 582.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyano-3-fluoropyridin-2-yl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 82

¹H NMR (400 MHz, DMSO-d₆): δ 8.36 (d, J=4.7 Hz, 1H), 7.70 (s, 1H), 7.39(m, 2H), 7.25-6.63 (m, 5H), 6.39 (s, 1H), 5.96 (s, 1H), 4.85 (s, 1H),4.34 (s, 1H), 3.12-2.69 (m, 3H), 2.64-2.01 (m, 5H). MS: 600.0 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyano-3-fluoropyridin-2-yl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 83

¹H NMR (400 MHz, DMSO-d₆): δ 8.37 (d, J=4.6 Hz, 1H), 7.75 (s, 1H), 7.39(m, 2H), 7.24-6.89 (m, 4H), 6.87-6.65 (d, 1H), 6.50-6.27 (m, 1H),5.59-5.40 (m, 1H), 4.92-4.75 (m, 1H), 4.05-3.87 (m, 1H), 2.95-2.68 (m,1H), 2.62-2.43 (m, 1H), 2.41-2.25 (m, 1H), 2.25-2.09 (m, 2H), 2.05-1.74(m, 4H), 1.59-1.24 (m, 3H). MS: 628.0 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyano-3-fluoropyridin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 88

¹H NMR (400 MHz, DMSO-d₆): δ 8.73 (s, 1H), 8.49 (m, 1H), 7.96 (s, 1H),7.59-7.30 (m, 3H), 7.26-6.68 (m, 6H), 6.52-6.12 (m, 1H), 5.96 (d, J=10.5Hz, 1H), 4.95 (s, 1H), 4.63 (m, 1H), 4.49 (m, 1H), 4.22 (s, 1H),4.14-4.02 (m, 1H), 3.46-2.65 (m, 4H), 2.55-2.00 (m, 2H), 1.69-1.49 (m,2H). MS: 618.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-1-(pyrazin-2-yl)pyrrolidine-2-carboxamide(Single Enantiomer)—Compound 58

¹H NMR (400 MHz, CDCl₃): δ 9.74 (d, J=1.5 Hz, 1H), 8.32 (m, 2H), 7.71(s, 1H), 7.36 (m, 1H), 7.16 (m, 1H), 6.97 (m, 4H), 6.41 (s, 1H), 5.44(d, J=7.0 Hz, 1H), 4.85 (d, J=6.0 Hz, 1H), 3.96 (m, 1H), 2.98-2.82 (m,1H), 2.61-2.48 (m, 1H), 2.35-2.21 (m, 1H), 2.02 (m, 5H), 1.88 (m, 2H),1.47-1.19 (m, 2H). MS: 586.2 (M+1)⁺.

2-(((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3-fluorophenyl)carbamoyl)-4-hydroxypyrrolidine-1-carboxylate (SingleEnantiomer)—Compound 74

¹H NMR (400 MHz, CDCl₃): δ 8.60 (s, 1H), 7.89 (s, 1H), 7.71 (s, 1H),7.45-7.29 (m, 2H), 7.25-6.86 (m, 5H), 6.41 (s, 1H), 5.54 (s, 1H), 4.98(s, 1H), 3.98 (s, 1H), 3.16-2.66 (m, 2H), 2.51 (s, 1H), 2.26 (s, 1H),1.98 (m, 7H), 1.55 (m, 3H). MS: 591.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluoro-phenyl)-2-oxo-3-(pyrimidin-2-yl)oxazolidine-4-carboxamide(Single Enantiomer)—Compound 76

¹H NMR (400 MHz, CDCl₃): δ 8.70 (d, J=4.7 Hz, 2H), 7.67 (d, J=8.0 Hz,1H), 7.43-7.31 (m, 1H), 7.19 (d, J=7.3 Hz, 1H), 7.13-6.86 (m, 5H), 6.46(s, 1H), 5.58 (d, J=6.8 Hz, 1H), 5.02 (d, J=4.4 Hz, 1H), 4.47 (dd,J=8.7, 5.0 Hz, 1H), 4.24-4.13 (m, 1H), 3.98 (s, 1H), 2.14-1.79 (m, 6H),1.57-1.41 (m, 2H). MS: 588.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-2-oxooxazolidine-4-carboxamide(Single Enantiomer)—Compound 77

¹H NMR (400 MHz, CDCl₃): δ 8.48 (s, 1H), 7.62 (d, J=7.9 Hz, 1H), 7.33(d, J=8.9 Hz, 1H), 7.19 (d, J=7.2 Hz, 2H), 7.10-6.85 (m, 5H), 6.44 (d,J=5.1 Hz, 1H), 6.20-6.08 (m, 1H), 5.01 (m, 1H), 4.46 (dd, J=8.7, 4.7 Hz,1H), 4.31-4.20 (m, 2H), 3.09-2.91 (m, 2H), 2.58-2.30 (m, 2H). MS: 584.1(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-3-(4-cyano-pyridin-2-yl)-N-(3-fluorophenyl)-2-oxooxazolidine-4-carboxamide(Single Enantiomer)—Compound 78

¹H NMR (400 MHz, CDCl₃): δ 8.55 (s, 1H), 8.50 (t, J=5.8 Hz, 1H), 7.67(d, J=8.5 Hz, 1H), 7.43-7.29 (m, 2H), 7.20 (d, J=7.6 Hz, 1H), 7.15-6.89(m, 4H), 6.43 (d, J=4.4 Hz, 1H), 5.54 (d, J=7.9 Hz, 1H), 5.06 (d, J=4.7Hz, 1H), 4.51 (dd, J=8.8, 5.0 Hz, 1H), 4.25 (m, 1H), 3.98 (s, 1H),2.19-1.74 (m, 6H), 1.49 (m, 2H). MS: 612.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-cyano-5-fluorophenyl)-3-(3-cyanophenyl)-2-oxooxazolidine-4-carboxamide(Single Enantiomer)—Compound 134

¹H NMR (400 MHz, CDCl₃): δ 8.51-8.47 (m, 1H), 8.39-8.37 (d, 0.5H),8.07-7.99 (m, 1H), 7.38 (s, 0.5H), 7.33-7.31 (m, 1H), 7.26-7.22 (m, 1H),7.08-7.07 (m, 1H), 6.90-6.87 (m, 1H), 6.53-6.46 (m, 2H), 4.94-4.91 (m,1H), 4.44-4.40 (m, 1H). 4.34-4.32 (m, 1H), 4.28-4.23 (m, 1H), 3.00-2.99(m, 2H), 2.50-2.43 (m, 2H). MS: 608.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-2-oxooxazolidine-4-carboxamide(Single Enantiomer)—Compound 135

¹H NMR (400 MHz, CDCl₃): δ 8.58-8.28 (m, 3H), 8.08 (d, J=8.5 Hz, 1H),7.32 (dd, J=5.1, 1.0 Hz, 2H), 7.28-7.20 (m, 1H), 7.07 (m, 1H), 6.91 (m,1H), 6.66-6.22 (m, 2H), 5.05-4.85 (m, 1H), 4.57-4.09 (m, 3H), 3.02 (m,2H), 2.69-2.30 (m, 2H). MS: 585.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-2-oxooxazolidine-4-carboxamide(Single Enantiomer)—Compound 132

¹H NMR (400 MHz, CDCl₃): δ 8.91 (s, 1H), 8.41 (m, 4H), 8.11 (s, 1H),7.23 (s, 1H), 7.05 (s, 1H), 6.91 (s, 1H), 6.52 (m, 1H), 6.05 (m, 1H),4.95 (m, 1H), 4.37 (m, 2H), 3.95 (s, 1H), 1.71 (m, 10H). MS: 613.2(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-2-oxo-3-(thiazol-4-yl)oxazolidine-4-carboxamide (SingleEnantiomer)—Compound 72

¹H NMR (400 MHz, CDCl₃): δ 8.70-8.47 (m, 1H), 7.69-7.52 (m, 1H), 7.49(d, J=2.0 Hz, 1H), 7.42-7.26 (m, 1H), 7.25-6.84 (m, 5H), 6.42 (s, 1H),6.21-6.02 (m, 1H), 5.03 (d, J=4.6 Hz, 1H), 4.42 (m, 1H), 4.38-4.05 (m,2H), 2.98 (m, 2H), 2.64-2.29 (m, 2H). MS: 565.1 (M+1)⁺.

(4S)—N-(1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluoro-5-(2-hydroxypropan-2-yl)phenyl)-2-oxooxazolidine-4-carboxamide(Racemic)—Compound 145

¹H NMR (400 MHz, CDCl₃): δ 8.63-8.50 (m, 1H), 8.42 (m, 1H), 7.48-7.40(m, 1H), 7.29 (d, J=7.0 Hz, 2H), 7.25-7.19 (m, 2H), 7.14-6.95 (m, 3H),6.89 (m, 1H), 6.67 (d, J=6.9 Hz, 1H), 6.54-6.42 (m, 1H), 5.11-4.96 (m,1H), 4.51-4.40 (m, 1H), 4.32 (d, J=9.1 Hz, 1H), 4.24-4.09 (m, 1H),3.12-2.73 (m, 2H), 1.52 (m, 2H), 1.32 (d, J=9.0 Hz, 4H). MS: 642.2(M+1)⁺.

(4S)—N-(1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-2-oxo-1,3-oxazinane-4-carboxamide(Racemic)—Compound 90

¹H NMR (400 MHz, CDCl₃): δ 8.57 (s, 1H), 8.40 (s, 1H), 7.68 (d, J=8.0Hz, 1H), 7.25-6.91 (m, 8H), 6.48 (s, 1H), 6.25 (s, 1H), 5.08 (s, 1H),4.51-4.46 (m, 1H), 4.31 (m, 2H), 3.01 (m, 2H), 2.53-2.50 (m, 2H),2.29-2.13 (m, 2H). MS: 598.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-2-oxo-1,3-oxazinane-4-carboxamide(Single Enantiomer)—Compound 133

¹H NMR (400 MHz, CDCl₃): δ 8.55 (d, J=5.0 Hz, 1H), 8.34 (s, 1H), 7.54(d, J=8.4 Hz, 1H), 7.31 (dd, J=5.0, 1.1 Hz, 1H), 7.26-7.16 (m, 2H),7.13-7.04 (m, 1H), 6.98 (t, J=6.6 Hz, 2H), 6.72-6.63 (m, 1H), 6.49 (s,1H), 6.44 (d, J=6.9 Hz, 1H), 5.11 (dd, J=6.4, 3.5 Hz, 1H), 4.51-4.22 (m,3H), 2.98-3.04 (m, 2H), 2.67-2.41 (m, 2H), 2.33-2.09 (m, 2H). MS: 627.2(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-2-oxo-1,3-oxazinane-4-carboxamide(Single Enantiomer)—Compound 139

¹H NMR (400 MHz, CDCl₃): δ 8.64 (d, J=5.0 Hz, 1H), 8.47 (s, 1H), 7.45(d, J=7.4 Hz, 1H), 7.38-7.30 (m, 2H), 7.24 (d, J=7.1 Hz, 1H), 7.15-7.12(m, 1H), 6.81-6.77 (m, 1H), 6.06 (s, 1H), 5.51 (d, J=7.5 Hz, 1H),5.05-4.88 (m, 1H), 4.62-4.56 (m, 1H), 4.42-4.30 (m, 1H), 3.87 (s, 1H),2.35-2.15 (m, 2H), 1.97-1.79 (m, 5H), 1.40 (m, 2H). MS: 643.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-3-(4-cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-2-oxo-1,3-oxazinane-4-carboxamide(Single Enantiomer)—Compound 144

¹H NMR (400 MHz, CDCl₃): δ 8.96 (d, J=4.7 Hz, 1H), 7.56 (d, J=10.0 Hz,1H), 7.41 (dd, J=9.7, 6.4 Hz, 2H), 7.24-7.22 (m, 1H), 7.14-6.95 (m, 3H),6.70 (t, J=8.6 Hz, 1H), 6.52 (s, 1H), 5.53 (d, J=7.6 Hz, 1H), 4.96 (dd,J=7.8, 4.0 Hz, 1H), 4.46 (d, J=8.8 Hz, 1H), 4.31 (dd, J=10.7, 5.1 Hz,1H), 3.99 (s, 1H), 2.49-2.31 (m, 1H), 2.29-2.01 (m, 5H), 1.98-1.78 (m,2H), 1.49 (dd, J=17.9, 8.5 Hz, 1H). MS: 645.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-3-(4-cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-2-oxo-1,3-oxazinane-4-carboxamide(Single Enantiomer)—Compound 154

¹H NMR (400 MHz, CDCl₃): δ 8.89 (d, J=4.8 Hz, 1H), 7.52 (d, J=8.9 Hz,1H), 7.40 (d, J=4.8 Hz, 1H), 7.22 (dd, J=8.0, 1.2 Hz, 1H), 7.16-7.15 (m,1H), 7.08-6.97 (m, 2H), 6.94 (dd, J=7.7, 1.5 Hz, 1H), 6.66 (dd, J=9.7,7.4 Hz, 1H), 6.56 (s, 1H), 6.43 (d, J=6.8 Hz, 1H), 4.91 (dd, J=8.3, 4.5Hz, 1H), 4.41-4.33 (m, 2H), 4.24-4.20 (m, 1H), 3.06-2.86 (m, 2H),2.66-2.42 (m, 2H), 2.39-2.25 (m, 1H), 2.24-2.12 (m, 1H). MS: 617.2(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-2-oxo-1,3-oxazinane-4-carboxamide(Single Enantiomer)—Compound 143

¹H NMR (400 MHz, CDCl₃): δ 9.08-7.79 (m, 3H), 7.62-6.70 (m, 5H), 6.50(m, 2H), 4.95 (m, 1H), 4.62-4.03 (m, 3H), 2.99 (s, 2H), 2.51 (s, 2H),2.18 (m, 2H). MS: 599.1 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-3-(4-cyano-pyridin-2-yl)-N-(5-fluoropyridin-3-yl)-2-oxo-1,3-oxazinane-4-carboxamide(Single Enantiomer)—Compound 137

¹H NMR (400 MHz, CDCl₃): δ 8.43-8.90 (m, 3H), 8.30 (s, 1H), 7.49-8.13(m, 1H), 7.29-7.31 (m, 2H), 7.17-7.21 (m, 1H), 6.94-7.08 (m, 2H),6.45-6.53 (m, 1H), 5.80-593 (m, 1H), 4.96-5.00 (m, 1H), 4.47-4.51 (m,1H), 4.30-4.33 (m, 1H), 3.96-3.98 (m, 1H), 2.09-2.28 (m, 6H), 1.83-1.95(m, 2H), 1.49-1.63 (m, 2H). MS: 627.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluoro-5-(2-hydroxypropan-2-yl)phenyl)-2-oxo-1,3-oxazinane-4-carboxamide(Single Enantiomer)—Compound 146

¹H NMR (400 MHz, CDCl₃): δ 8.56 (t, J=6.0 Hz, 1H), 8.36 (s, 1H),7.72-7.45 (m, 1H), 7.23-7.16 (m, 1H), 7.12 (t, J=7.1 Hz, 1H), 7.06-6.86(m, 3H), 6.38 (s, 1H), 6.28 (d, J=6.9 Hz, 1H), 5.17-5.01 (m, 1H),4.50-4.44 (m, 1H), 4.30 (m, 2H), 2.99 (d, J=7.8 Hz, 2H), 2.62-2.37 (m,2H), 2.36-2.06 (m, 2H), 1.49 (d, J=6.2 Hz, 2H), 1.32 (m, 4H). MS: 656.2(M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-6-oxopiperidine-2-carboxamide(Single Enantiomer)—Compound 55

¹H NMR (400 MHz, CDCl₃): δ 8.59 (s, 1H), 8.28 (s, 1H), 7.72 (d, J=7.2Hz, 1H), 7.43-7.33 (m, 1H) 7.26-7.12 (m, 2H), 7.11-6.96 (m, 2H), 6.89(dd, J=8.3, 2.2 Hz, 1H), 6.46 (s, 1H), 6.27 (s, 1H), 5.00 (t, J=4.6 Hz,1H), 4.37-4.28 (m, 1H), 3.13-2.95 (m, 2H), 2.78-2.69 (m, 1H), 2.62-2.35(m, 3H), 2.15-2.09 (m, 1H), 2.05-1.92 (m, 1H), 1.89-1.70 (m, 3H). MS:596.2 (M+1)⁺.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyano-pyridin-2-yl)-N-(3-fluorophenyl)-6-oxopiperidine-2-carboxamide(Single Enantiomer)—Compound 75

¹H NMR (400 MHz, CDCl₃): δ 8.60 (s, 1H), 8.31 (s, 1H), 7.73-7.75 (m,1H), 7.30 (m, 1H), 7.00-7.17 (m, 5H), 6.87-6.91 (m, 1H), 6.45 (s, 1H),5.50 (d, J=7.0 Hz, 1H), 5.00-5.02 (m, 1H), 3.99 (m, 1H), 2.60-2.74 (m,1H), 2.58-2.60 (m, 1H), 2.01-2.14 (m, 6H), 1.83-1.92 (m, 4H), 1.42-1.46(m, 3H). MS: 624.2 (M+1)⁺.

(2S,4R)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-4-fluoro-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 151

¹H NMR (400 MHz, CDCl₃): δ 8.75 (s, 1H), 8.57 (s, 1H), 7.76 (s, 1H),7.36 (m, 2H), 7.06 (m, 6H), 6.39 (s, 1H), 5.51 (d, J=6.6 Hz, 1H), 5.12(m, 1H), 4.82 (s, 1H), 3.91 (m, 1H), 2.69-2.26 (m, 2H), 2.05 (m, 6H),1.53-1.38 (m, 2H). MS: 628.2 (M+1)⁺.

Example 9 Preparation of(2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-dicyanophenyl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 191

Step A: 5-Nitroisophthaloyl dichloride

To a solution of 5-nitroisophthalic acid (2.3 g, 11 mmol) in SOCl₂ (6mL) was added a drop of DMF and the mixture was stirred at reflux for 3hr. The resulting reaction mixture was concentrated to give the crudeproduct which was used directly in the next step.

Step B: 5-Nitroisophthalamide

5-Nitroisophthaloyl dichloride (2.7 g, 9.7 mmol) was added portionwiseto a cold solution of NH₃.H₂O (40 mL) at 0° C. The reaction mixture wasstirred overnight and a white precipitate formed. The mixture was thenfiltered, washed with excess of water, and dried at 110° C. to give thecrude product which was used directly in the next step.

Step C: 5-Aminoisophthalamide

To a solution of 5-nitroisophthalamide (2 g, 9.6 mmol) in MeOH (200 mL)was added Pd/C (200 mg). The reaction was stirred overnight under ahydrogen atmosphere. The suspension was filtered and the filtrate wasconcentrated to afford the desired product which was used directly inthe next step.

Step D:5-((2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)1-(4cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamido)isophthalamide

A mixture of 2-chlorobenzaldehyde (1.0 mL, 7.3 mmol) and5-aminoisophthalamide (1.3 g, 7.3 mmol) was stirred at room temperaturefor 30 min under N₂, followed by addition of(S)-1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxylic acid (1.7 g,7.3 mmol). After stirring for 10 min, 1,1-difluoro-3-isocyanocyclobutane(854 mg, 7.3 mmol) was added. The mixture was then stirred overnight andfiltered and purified by a standard method to give the title product.

Step E:(2S)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)1-(4-cyanopyridin-2-yl)-N-(3,5-dicyanophenyl)-5-oxopyrrolidine-2-carboxamide

To a mixture of5-((2S)—N-(1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamido)isophthalamide (850 mg, 1.3mmol) in pyridine (0.62 mL, 7.8 mmol) and DCM (10 mL) was added TFAA(0.9 mL, 6.5 mmol). The reaction solution was stirred at roomtemperature overnight. The resulting mixture was concentrated and theresidue was purified by a standard method to afford the title product.¹H NMR (400 MHz, CDCl₃): δ 8.77 (s, 1H), 8.62-8.42 (m, 2H), 7.87 (s,1H), 7.75 (s, 1H), 7.40 (d, J=7.8 Hz, 1H), 7.31 (d, J=4.2 Hz, 1H), 7.25(d, J=8.1 Hz, 1H), 7.10 (t, J=7.3 Hz, 1H), 6.92 (d, J=7.5 Hz, 1H), 6.47(s, 1H), 6.11 (d, J=6.6 Hz, 1H), 4.73 (dd, J=9.4, 2.7 Hz, 1H), 4.35 (s,1H), 3.14-2.82 (m, 3H), 2.68-2.31 (m, 3H), 2.19 (m, 1H), 2.09-1.91 (m,1H). MS: 614.1 (M+1)⁺.

The following analogs were synthesized via the procedure set forthabove, using the appropriate aldehyde, amine, carboxylic acid,isocyanide and halo-substituted aromatic ring or heterocyclic(heteroaromatic) ring using the reagents and solvents set forth above,and purified via standard methods.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-dicyanophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 153

¹H NMR (400 MHz, CDCl₃): δ 8.74 (s, 1H), 8.53 (m, 2H), 7.81 (m, 2H),7.48-7.16 (m, 4H), 7.09 (t, J=7.5 Hz, 1H), 6.90 (d, J=7.6 Hz, 1H), 6.46(s, 1H), 6.17 (d, J=6.7 Hz, 1H), 4.72 (dd, J=9.1, 2.3 Hz, 1H), 4.35 (s,1H), 3.18-2.71 (m, 3H), 2.68-1.83 (m, 5H). MS: 614.1 (M+1)⁺.

Example 10 Preparation of (S)-tert-butyl3-(((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-5-oxopiperazine-1-carboxylate(Single Enantiomer)—Compound 97

Compound 97 was synthesized via the UGI reaction procedure set forthherein, using the appropriate aldehyde, amine, carboxylic acid,isocyanide and halo-substituted aromatic ring or heterocyclic(heteroaromatic) ring and purified via standard methods.

¹H NMR (400 MHz, CDCl₃): δ 8.75-8.44 (m, 2H), 7.81-7.41 (m, 1H),7.46-7.35 (m, 2H), 7.24 (t, J=7.2 Hz, 1H), 7.16-6.97 (m, 2H), 6.84-6.75(m, 2H), 6.43-5.82 (m, 1H), 5.09-4.98 (m, 1H), 4.77-4.73 (m, 1H), 4.48(d, J=13.5 Hz, 1H), 4.27-4.07 (m, 2H), 3.45-2.76 (m, 4H), 1.54 (s, 9H).MS: 613.2 (M+1)⁺.

Example 11 Preparation of (3S)-tert-butyl3-((1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4-cyanopyrimidin-2-yl)-5-oxopiperazine-1-carboxylate(Racemic)—Compound 98

A mixture of(3S)-tert-butyl3-((1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-5-oxopiperazine-1-carboxylate (200 mg,0.326 mmol), 2-bromopyrimidine-4-carbonitrile (0.489 mmol), Pd₂(dba)₃(30.2 mg, 0.0323 mmol), XantPhos (19.1 mg, 0.03 mmol) and Cs₂CO₃ (148.7mg, 0.46 mmol) in 1,4-dioxane (10 mL) was stirred at 80° C. for 3 hrunder N₂. The reaction mixture was cooled to room temperature andfiltered. The filtrate was concentrated and the residue was purified bya standard method to afford the desired product. ¹H NMR (400 MHz,CDCl₃): δ 8.97 (d, J=4.3 Hz, 1H), 7.85-7.55 (d, 1H), 7.51-7.39 (m, 2H),7.25 (t, J=7.6 Hz, 1H), 7.13-6.26 (m, 6H), 5.91 (d, J=7.6 Hz, 1H),4.92-4.08 (m, 5H), 3.38 (t, J=14.9 Hz, 1H), 3.02 (s, 2H), 2.83-2.22 (d,2H), 1.61 (s, 9H). MS: 716.1 (M+1)⁺.

The following analogs were synthesized via the procedure set forthabove, using the appropriate aldehyde, amine, carboxylic acid, isocynideand halo-substituted aromatic ring or heterocyclic (heteroaromatic) ringusing the reagents and solvents set forth above, and purified viastandard methods.

(S)-tert-Butyl3-4(S)-1-(2-chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4-cyanopyrimidin-2-yl)-5-oxopiperazine-1-carboxylate(Chiral)—Compound 93

¹H NMR (400 MHz, CDCl₃): δ 8.96 (d, J=4.3 Hz, 1H), 7.83 (s, 1H), 7.43(m, 2H), 7.21 (t, J=7.4 Hz, 1H), 7.08-6.62 (m, 4H), 6.63-6.37 (m, 1H),5.93 (m, 1H), 4.85 (t, J=3.6 Hz, 1H), 4.63-4.23 (m, 2H), 4.16 (m, 1H),3.93 (s, 1H), 3.43 (m, 1H), 2.24-1.91 (m, 5H), 1.79 (m, 3H), 1.60 (m,1H). MS: 744.2 (M+1)⁺.

(3S)-tert-Butyl3-((1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3-fluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)-5-oxopiperazine-1-carboxylate(Single Enantiomer)—Compound 89

¹H NMR (400 MHz, DMSO-d₆): δ 8.80-8.37 (m, 1H), 8.05-7.57 (m, 1H),7.58-7.31 (m, 3H), 7.21 (s, 1H), 7.16-6.89 (m, 3H), 6.90-6.68 (m, 1H),6.67-6.30 (m, 1H), 6.22-5.84 (m, 1H), 5.09-4.87 (m, 1H), 5.83-4.57 (m,1H), 4.50 (m, 1H), 4.25 (s, 1H), 4.08 (m, 1H), 3.50-2.70 (m, 4H),2.60-2.10 (m, 1H), 1.70 (s, 2H), 1.54 (m, 1H). MS: 697.2 (M+1)⁺.

Example 12 Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-6-oxopiperazine-2-carboxamide(Single Enantiomer)—Compound 99

TFA (0.3 mL) was added to a solution of (S)-tert-butyl3-(((R)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)-5-oxopiperazine-1-carboxylate(60 mg, 0.08 mmol) in DCM (1.0 mL) at 0° C. The mixture was warmed toroom temperature and stirred for 1 hr, and then concentrated. Theresidue was purified by a standard method to give the desired product.¹H NMR (400 MHz, CDCl₃): δ 8.94 (t, J=4.6 Hz, 1H), 7.48-7.36 (m, 3H),7.21 (m, J=7.8, 1.5 Hz, 1H), 7.12-6.94 (m, 3H), 6.71-6.55 (m, 2H), 6.05(d, J=6.7 Hz, 1H), 4.73 (q, J=4.0, 2.1 Hz, 1H), 4.36 (d, J=6.7 Hz, 1H),3.77-3.65 (m, 2H), 3.50-3.35 (m, 1H), 3.18 (m, 1H), 3.12-2.96 (m, 2H),2.64-2.35 (m, 2H). MS: 616.1 (M+1)⁺.

The following compound was synthesized via the procedure set forthabove, using the appropriate aldehyde, amine, carboxylic acid,isocyanide and halo-substituted aromatic ring or heterocyclic(heteroaromatic) ring using the reagents and solvents set forth above,and purified via standard methods.

(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-6-oxopiperazine-2-carboxamide(Single Enantiomer)—Compound 100

¹H NMR (400 MHz, CDCl₃): δ 8.68-8.28 (m, 1H), 7.61-7.28 (m, 2H), 7.20(dd, J=7.9, 1.3 Hz, 0H), 7.02-6.90 (m, 1H), 6.66 (tt, J=8.6, 2.3 Hz,1H), 6.49 (d, J=2.7 Hz, 0H), 6.09 (m, 1H), 4.90 (dd, J=3.8, 2.0 Hz, 1H),4.42-4.16 (m, 1H), 3.71 (m, 1H), 3.50-3.23 (m, 1H), 3.18-2.78 (m, 2H),2.63-2.13 (m, 2H). MS: 615.2 (M+1)⁺.

Example 13(S)-4-Acetyl-N—((S)-1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-6-oxopiperazine-2-carboxamide(Single Enantiomer)—Compound 92

To a solution of (3S)-tert-butyl3-((1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)(3-fluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)-5-oxopiperazine-1-carboxylate(100 mg, 0.14 mmol) in DCM (3 mL) was added TFA dropwise (1 mL) at 0° C.The mixture was stirred at room temperature for 2 hr and thenconcentrated. The residue was dissolved in DCM and cooled to 0° C. DIPEA(0.055 mL, 0.34 mmol) was added to the mixture followed by Ac₂O (0.031mL, 0.34 mmol) at 0° C. Then the mixture was stirred at room temperaturefor 2 hr. The solution was concentrated and the residue was purified bya standard method to afford the desired product. ¹H NMR (400 MHz,CDCl₃): δ 8.54 (s, 2H), 7.70-744 (m, 2H), 7.36 (m, 2H), 7.20 (t, J=7.2Hz, 1H), 7.14-6.99 (m, 2H), 6.94 (t, J=7.4 Hz, 1H), 6.80 (s, 1H), 6.66(d, J=7.8 Hz, 1H), 6.58-6.42 (m, 1H), 5.09 (dt, J=5.2, 3.1 Hz, 1H), 4.93(m, 1H), 4.63 (m, 1H), 4.54-4.41 (m, 1H), 4.35-4.31 (m, 1H), 3.16 (s,1H), 3.12-2.96 (m, 3H), 2.86 (s, 1H), 2.25 (s, 3H). MS: 639.2 (M+1)⁺.

Example 14 Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-4-cyclopropyl-N-(3,5-difluorophenyl)-6-oxopiperazine-2-carboxamide(Single Enantiomer)—Compound 106

TFA (0.3 mL) was added to a solution of (S)-tert-butyl3-(((R)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)-5-oxopiperazine-1-carboxylate(60 mg, 0.084 mmol) in DCM (1.0 mL) at 0° C. The mixture was stirred atroom temperature for 1 hr then concentrated. The residue was dissolvedin MeOH (2 mL) followed by addition of(1-ethoxycyclopropoxy)trimethylsilane (88 mg, 0.50 mmol), AcOH (50 mg,0.84 mmol) and NaBH₃(CN) (27 mg, 0.42 mmol). The resulting suspensionwas stirred at 80° C. under N₂ for 1.5 hr. The reaction mixture waspartitioned between EtOAc and H₂O. The organic layer was separated,washed with brine, dried over anhydrous Na₂SO₄ and concentrated. Theresidue was purified by a standard method to afford the desired product.¹H NMR (400 MHz, CDCl₃): δ 7.64 (d, J=7.8 Hz, 1H), 7.30 (d, J=5.3 Hz,2H), 7.19 (s, 1H), 7.07 (s, 3H), 6.66 (s, 1H), 6.32 (s, 1H), 6.09 (m,1H), 5.09 (s, 1H), 4.28 (s, 1H), 3.76-3.59 (m, 1H), 3.46-3.33 (m, 1H),3.08-2.89 (m, 4H), 2.59-2.31 (m, 2H), 0.94 (s, 1H), 0.61-0.37 (m, 4H).MS: 655.2 (M+1)⁺.

Example 15 Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4-methyl-6-oxopiperazine-2-carboxamide(Single Enantiomer)—Compound 101

TFA (0.6 mL) was added to a solution of (3S)-tert-butyl3-((1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)-5-oxopiperazine-1-carboxylate(30 mg, 0.042 mmol) in DCM (2 mL) at 0° C. The mixture was stirred atroom temperature for 1 hr and then concentrated. The residue wasdissolved in MeCN (4 mL) followed by addition of K₂CO₃ (10 mg, 0.072mmol) and iodomethane (2 mL). The resulting mixture was stirred at roomtemperature for 2 hr and then concentrated. The residue was purified bya standard method to afford the desired product. ¹H NMR (400 MHz,CDCl3): δ 8.60 (m, 2H), 7.80 (s, 1H), 7.45 (d, J=7.9 Hz, 1H), 7.33 (m,1H), 7.07 (d, J=4.3 Hz, 2H), 6.74 (t, J=8.6 Hz, 1H), 6.48-5.91 (m, 3H),4.92 (t, J=4.7 Hz, 1H), 4.20 (m, 1H), 3.61-3.40 (m, 1H), 3.14 (m, 1H),3.02-2.77 (m, 3H), 2.71 (m, 1H), 2.42-2.26 (m, 5H), 2.04 (d, J=9.0 Hz,1H). MS: 629 (M+1)⁺.

Example 16 Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4-(2-hydroxyethyl)-6-oxopiperazine-2-carboxamide(Single Enantiomer)—Compound 107

To a solution of (S)-tert-butyl3-(((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)-5-oxopiperazine-1-carboxylate(30 mg, 0.04 mmol) in DCM (3 mL) was added TFA (1 mL) at 0° C. Themixture was stirred at room temperature for 1 hr and concentrated invacuo. The residue was dissolved in EtOH (3 mL) followed by addition ofTBAI (16 mg, 0.04 mmol), Et₃N (10 mg, 0.1 mol) and 2-bromoethanol (7 mg,0.056 mmol). The resulting mixture was stirred at 85° C. for 3 hr andthen filtered. The filtrate was concentrated and the residue waspurified by a standard method to afford the desired product. ¹H NMR (400MHz, CDCl₃): δ 8.96 (t, J=4.6 Hz, 1H), 7.53-7.36 (m, 3H), 7.23 (m,J=7.8, 1.5 Hz, 1H), 7.14-6.94 (m, 3H), 6.68 (m, J=8.6, 2.3 Hz, 1H), 6.60(d, J=3.1 Hz, 1H), 6.07 (d, J=6.7 Hz, 1H), 4.75 (q, J=4.0, 2.1 Hz, 1H),4.38 (d, J=6.7 Hz, 1H), 3.78-3.67 (m, 2H), 3.39 (m, 1H), 3.26-2.92 (m,3H), 2.67-2.36 (m, 2H). MS: 659.2 (M+1)⁺.

The following compound was synthesized via the procedure set forthabove, using the appropriate aldehyde, amine, carboxylic acid,isocyanide and halo-substituted aromatic ring or heterocyclic(heteroaromatic) ring using the reagents and solvents set forth above,and purified via standard methods.

¹H NMR (400 MHz, CDCl₃): δ 8.60-8.56 (m, 2H), 7.47-7.28 (m, 3H),7.22-7.01 (m, 4H), 6.72-6.67 (m, 1H), 6.54-6.44 (m, 2H), 5.24 (m, 1H),4.37-4.13 (m, 3H), 3.63-2.97 (m, 8H), 2.44-2.06 (m, 2H), 1.34-1.28 (m,3H). MS: 701.2 (M+1)⁺.

Example 17 Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(5-cyanooxazol-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 162

Step A: Oxazole-5-carboxamide

Ethyl oxazole-5-carboxylate (2 g, 14.2 mmol) was dissolved in NH₃solution (7 M in MeOH, 25 mL). The solution was stirred at roomtemperature for 2 hr and filtered. The solid was dried to give thedesired product (1.5 g, 92% yield) as a white powder which was useddirectly in the next step.

Step B: 2-Iodooxazole-5-carboxamide

Oxazole-5-carboxamide (560 mg, 5.0 mmol) was dissolved in anhydrous THF(7.5 mL) and flushed with N₂. The solution was cooled to −42° C. andtreated with fresh LiHMDS (15 mL, 1 M in THF). The solution became darkyellow was stirred for 20 min and followed by the addition of a solutionof ZnCl₂ (30 mL, 0.5 M in THF). The reaction was warmed to 0° C. for 1hr. After solid iodine (1.65 g, 6.5 mmol) was added, the reactionmixture was stirred at room temperature for another 1 hr and then pouredinto saturated Na₂S₂O₃ solution containing 25% aq. NH₃ solution. Theresulting mixture was extracted with EtOAc (3×30 mL). The combinedorganic layers were washed with brine, dried over anhydrous Na₂SO₄ andconcentrated. The resulting residue was purified by a standard method togive the desired product. MS: 239.0 (M+1)⁺.

Step C:2-((S)-2-(((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-5-oxopyrrolidin-1-yl)oxazole-5-carboxamide

The product was prepared by the general procedure for the Buchwaldreaction. ¹H NMR (400 MHz, CDCl₃): δ 7.59 (s, 1H), 7.53 (s, 1H), 7.37(d, J=7.9 Hz, 1H), 7.20 (t, J=7.0 Hz, 1H), 7.04 (t, J=7.6 Hz, 1H), 6.96(d, J=7.9 Hz, 2H), 6.68 (t, J=8.7 Hz, 1H), 6.46 (s, 1H), 6.36 (d, J=6.4Hz, 1H), 5.68 (s, 1H), 4.82 (dd, J=9.3, 2.6 Hz, 1H), 4.33 (s, 1H),4.16-4.09 (m, 1H), 3.03-3.00 (m, 2H), 2.90-2.77 (m, 1H), 2.62-2.35 (m,3H), 2.29-2.28 (m, 1H), 2.19-2.08 (m, 1H). MS: 608.1 (M+1)⁺.

Step D:(S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(5-cyanooxazol-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide

2-((S)-2-(((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluoro-phenyl)carbamoyl)-5-oxopyrrolidin-1-yl)oxazole-5-carboxamide(100 mg, 0.16 mmol) was dissolved in DCM (3 mL) and dry pyridine (0.8mL). TFAA (0.1 mL) was added and the reaction solution was stirred for25 min at room temperature and then concentrated in vacuo. The residuewas dissolved in EtOAc and washed with H₂O, saturated aq. NaHCO₃ andbrine. The organic phase was separated, dried over anhydrous Na₂SO₄, andconcentrated. The residue was purified by a standard method to give thedesired product. ¹H NMR (400 MHz, CDCl₃): δ 7.63 (s, 1H), 7.55 (d, J=7.0Hz, 1H), 7.41 (d, J=7.1 Hz, 1H), 7.25 (td, J=7.8, 1.5 Hz, 1H), 7.08 (t,J=7.6 Hz, 1H), 6.98-6.91 (m, 1H), 6.80 (d, J=6.7 Hz, 1H), 6.71 (dd,J=9.7, 7.4 Hz, 1H), 6.49 (s, 1H), 5.97 (d, J=6.8 Hz, 1H), 4.80 (dd,J=9.3, 2.8 Hz, 1H), 4.36 (s, 1H), 3.06-3.03 (m, 2H), 2.92-2.79 (m, 1H),2.62-2.29 (m, 4H), 2.18-2.12 (m, 1H). MS: 590.1 (M+1)⁺.

Example 18 Preparation of(2S,4R)—N-(1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-cyano-phenyl)-1-(4-cyanopyridin-2-yl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 170

Step A: (2S,4R)-1-tert-Butyl 2-methyl4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate

Imidazole (2.8 g, 40.8 mmol) was added to a solution of(2S,4R)-1-tert-butyl 2-methyl 4-hydroxypyrrolidine-1,2-dicarboxylate(5.0 g, 20.4 mmol) and TBSCl (4.6 g, 30.6 mmol) in anhydrous DMF (100mL). The mixture was stirred at room temperature overnight and thenpartitioned between EtOAc and H₂O. The organic layer was separated,washed with aq. LiCl (10%) and brine, dried over anhydrous Na₂SO₄, andthen concentrated. The residue was purified by column chromatography toafford the desired product as a colorless oil. MS: 360.2 (M+1)⁺.

Step B: (2S,4R)-1-tert-Butyl 2-methyl4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-1,2-dicarboxylate

To a solution of NaIO₄ (7.5 g, 35.0 mmol) in water (80 mL) was addedRuO₂ (370 mg, 2.8 mmol) under the atmosphere of nitrogen. The resultinggreen-yellow solution was stirred for 5 min followed by addition of(2S,4R)-1-tert-butyl-2-methyl4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate(5.0 g, 14.0 mmol) in EtOAc (44 mL) in one portion. The mixture wasstirred at room temperature overnight. The resulting mixture was thendiluted with EtOAc and filtered through a pad of Celite. The organiclayer was separated and washed with saturated aq. NaHSO₃, which resultedin precipitation of Ru black. The organic layer was then washed withbrine and dried over anhydrous Na₂SO₄. Evaporation of the solvent gavethe desired product as a colorless oil. MS: 374.2 (M+1)⁺.

Step C:(2S,4R)-4-((tert-Butyldimethylsilyl)oxy)-5-oxopyrrolidine-2-carboxylicacid

TFA (6 mL) was added to a solution of (2S,4R)-1-tert-butyl 2-methyl4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-1,2-dicarboxylate (2.5g, 6.68 mmol) in DCM (18 mL) at 0° C. The mixture was stirred at roomtemperature for 1 h then concentrated. The residue was dissolved inMeOH/THF (10 mL/10 mL) followed by addition of a solution of LiOH (842mg, 20.1 mmol) in water (5 mL). The resulting mixture was stirred atroom temperature for 1 h and then partitioned between EtOAc and H₂O. Theaqueous layer was separated and then adjusted to pH=6 with 1 N HCl aq.and extracted with EtOAc (3×20 mL). Combined organic layers were washedwith brine, dried over anhydrous Na₂SO₄, and then concentrated to affordthe desired product. ¹H NMR (400 MHz, DMSO-d₆): δ 12.87 (s, 1H), 8.17(s, 1H), 4.21 (t, J=8.0 Hz, 1H), 4.02 (d, J=8.4 Hz, 1H), 2.39-2.23 (m,1H), 2.09 (m, 1H), 0.84 (s, 9H), 0.07 (s, 6H). MS: 260.1 (M+1)⁺.

Step D

The same as general procedure for UGI reaction set forth herein.

Step E

The same as general procedure for Buchwald reaction set forth herein.

Step F:(2S,4R)—N-(1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-cyano-phenyl)-1-(4-cyanopyridin-2-yl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide

TBAF in THF (1N, 0.3 mL) was added to a solution of(2S,4R)-4-((tert-butyldimethylsilyl)oxy)-N-(1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-cyanophenyl)-1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide (0.15 mmol) in THF at 0° C.and the reaction solution was stirred at this temperature for 20 min.The resulting mixture was concentrated and the residue was purified by astandard method to afford the desired product. ¹H NMR (400 MHz, CDCl₃):δ 8.82-8.43 (m, 2H), 8.40-8.17 (m, 1H), 7.63-7.30 (m, 3H), 7.26-6.66 (m,4H), 6.68-6.34 (m, 2H), 6.65-6.31 (m, 2H), 4.87-4.56 (m, 2H), 4.23 (m,1H), 4.01-3.76 (m, 1H), 3.15-1.96 (m, 6H). MS: 605.1 (M+1)⁺.

The following analogs were synthesized via the procedure set forthherein, using the appropriate aldehyde, amine, carboxylic acid,isocynide and halo-substituted aromatic ring or heterocyclic(heteroaromatic) ring using the reagents and solvents set forth herein,and purified via various standard methods.

(2S,4R)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyano-pyridin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 113

¹H NMR (400 MHz, CDCl₃): δ 8.70 (m, 1H), 8.53 (s, 1H), 7.72 (d, J=7.5Hz, 1H), 7.32 (d, J=4.9 Hz, 2H), 7.18 (d, J=6.0 Hz, 1H), 7.09-6.85 (m,4H), 6.43 (s, 1H), 6.20 (d, J=5.3 Hz, 1H), 4.89 (s, 1H), 4.74 (t, J=9.2Hz, 1H), 4.37-4.32 (m, 1H), 3.40 (s, 1H), 3.11-2.87 (m, 2H), 2.77-2.14(m, 3H), 2.03-1.91 (m, 1H). MS: 598.1 (M+1)⁺.

(2S,4R)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide—Compound120

¹H NMR (400 MHz, CDCl₃): δ 8.98 (d, J=4.4 Hz, 1H), 7.70 (s, 1H), 7.39(d, J=4.9 Hz, 2H), 7.20-6.86 (m, 4H), 6.50 (s, 1H), 5.75 (s, 1H), 5.35(s, 1H), 4.92-4.63 (m, 2H), 4.34 (s, 1H), 2.91 (m, 3H), 2.21 (m, 4H).MS: 599.1 (M+1)⁺.

(2S,4R)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 121

¹H NMR (400 MHz, CDCl₃): δ 8.78 (s, 1H), 8.54 (s, 1H), 7.77 (d, J=8.1Hz, 1H), 7.45-7.30 (m, 2H), 7.25-6.83 (m, 5H), 6.42 (s, 1H), 5.49 (d,J=7.4 Hz, 1H), 4.83 (m 2H), 4.00 (s, 1H), 3.02 (s, 1H), 2.74 (m, 1H),2.25-1.74 (m, 7H), 1.56-1.33 (m, 2H). MS: 626.2 (M+1)⁺.

(2S,4R)—N—((R)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 122

¹H NMR (400 MHz, CDCl₃): δ 9.00 (d, J=4.8 Hz, 1H), 7.83 (m, 1H), 7.42(t, J=6.6 Hz, 2H), 7.22 (m, 2H), 7.18-7.08 (m, 1H), 7.08-6.67 (m, 2H),6.17 (m, 1H), 5.70 (d, J=7.6 Hz, 1H), 4.93-4.66 (m, 2H), 3.88 (d, J=7.7Hz, 1H), 3.37 (s, 1H), 2.71 (m, 1H), 2.03 (m, 5H), 1.88-1.64 (m, 4H).MS: 627.2 (M+1)⁺.

(2S,4R)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide—Compound123

¹H NMR (400 MHz, CDCl₃): δ 8.99 (d, J=4.4 Hz, 1H), 7.74 (d, J=7.9 Hz,1H), 7.47-7.29 (m, 3H), 7.08 (m, 6H), 6.51 (s, 1H), 5.61 (s, 1H), 4.81(m, 2H), 4.02 (d, J=7.2 Hz, 1H), 3.38 (s, 1H), 2.89-2.65 (m, 1H),2.23-1.81 (m, 8H), 1.58-1.48 (m, 1H). MS: 627.2 (M+1)⁺.

(2S,4R)—N—((R)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 114

¹H NMR (400 MHz, CDCl₃): δ 8.71 (d, J=5.8 Hz, 1H), 8.64-8.50 (m, 1H),7.94-7.56 (m, 1H), 7.47-7.31 (m, 2H), 7.29 (d, J=2.2 Hz, 1H), 7.26-7.18(m, 1H), 7.16-6.95 (m, 2H), 6.88-6.65 (m, 1H), 6.44-6.35 (m, 1H), 6.29(s, 1H), 6.11 (d, J=6.7 Hz, 1H), 4.77 (m, 2H), 4.40-4.08 (m, 1H), 3.27(s, 1H), 3.09-2.58 (m, 3H), 2.54-2.12 (m, 2H), 2.10-1.95 (m, 1H). MS:616 (M+1)⁺.

(2S,4R)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 115

1H NMR (400 MHz, MeOD): δ 8.65-8.50 (m, 2H), 7.54 (d, J=9.5 Hz, 1H),7.43-7.32 (m, 1H), 7.22-7.12 (m, 2H), 7.03 (m, 1H), 6.97-6.87 (m, 1H),6.84-6.75 (m, 2H), 6.36 (d, J=8.5 Hz, 1H), 4.89 (d, J=8.6 Hz, 1H),4.65-4.49 (m, 2H), 4.13 (m, 1H), 2.93-2.72 (m, 2H), 2.57-2.26 (m, 3H),1.85 (m, 1H). MS: 616.1 (M+1)⁺.

(2S,4R)—N—((R)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 116

¹H NMR (400 MHz, CDCl₃): δ 8.98 (t, J=5.0 Hz, 1H), 7.88 (s, 1H), 7.88(s, 1H), 7.50-7.37 (m, 2H), 7.33-7.20 (m, 2H), 7.19-7.06 (m, 2H),6.83-6.66 (m, 1H), 6.48 (m, 2H), 6.27 (s, 1H), 4.23 (s, 1H), 3.32 (s,1H), 2.87 (m, 2H), 2.66 (m, 1H), 2.35-2.02 (m, 3H). MS: 617.1 (M+1)⁺.

(2S,4R)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide—Compound117

¹H NMR (400 MHz, MeOD): δ 8.88 (d, J=4.9 Hz, 1H), 7.56 (m, 2H), 7.34(dd, J=8.0, 1.1 Hz, 1H), 7.16 (td, J=7.8, 1.6 Hz, 1H), 7.09-7.00 (m,1H), 6.98-6.85 (m, 2H), 6.81 (m, 1H), 6.42 (s, 1H), 4.87 (d, J=8.8 Hz,1H), 4.59-4.42 (m, 2H), 4.27-4.09 (m, 1H), 2.98-2.74 (m, 2H), 2.46 (m,3H), 2.02-1.76 (m, 1H). MS: 617.1 (M+1)⁺.

(2S,4R)—N—((R)-1-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 124

¹H NMR (400 MHz, CDCl₃): δ 8.71 (s, 1H), 8.64 (d, J=5.0 Hz, 1H), 7.79(s, 1H), 7.45 (d, J=7.8 Hz, 1H), 7.35 (dd, J=5.0, 1.0 Hz, 1H), 7.30-7.24(m, 1H), 7.16 (d, J=6.3 Hz, 1H), 7.14-7.05 (m, 1H), 6.79-6.68 (m, 2H),6.27 (s, 1H), 5.87 (d, J=7.5 Hz, 1H), 4.82 (t, J=6.9 Hz, 1H), 4.74 (t,J=9.2 Hz, 1H), 3.90-3.71 (m, 1H), 3.27 (s, 1H), 2.65 (m, 1H), 2.15-1.72(m, 8H), 1.57-1.43 (m, 1H). MS: 644.2 (M+1)⁺.

(2S,4R)—N—((S)-1-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 125

¹H NMR (400 MHz, CDCl₃): δ 8.83-8.47 (m, 2H), 7.62 (d, J=8.0 Hz, 1H),7.38 (d, J=8.0 Hz, 1H), 7.32 (d, J=5.0 Hz, 1H), 7.21 (t, J=7.1 Hz, 1H),7.05 (t, J=7.5 Hz, 1H), 6.98 (d, J=7.7 Hz, 1H), 6.85 (d, J=7.7 Hz, 1H),6.68 (t, J=8.6 Hz, 1H), 6.40 (s, 1H), 5.62 (d, J=7.7 Hz, 1H), 4.96-4.70(m, 2H), 4.01 (d, J=7.6 Hz, 1H), 3.37 (s, 1H), 2.70 (m, 1H), 2.14-1.74(m, 8H), 155-1.41 (m, 1H). MS: 644.2 (M+1)⁺.

(2S,4R)—N—((R)-1-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 126

¹H NMR (400 MHz, CDCl₃): δ 8.98 (dd, J=4.7, 2.1 Hz, 1H), 7.63 (d, J=7.3Hz, 1H), 7.50-7.33 (m, 2H), 7.28-6.87 (m, 3H), 6.84-6.38 (m, 2H), 6.19(s, 1H), 5.82 (d, J=7.6 Hz, 1H), 4.94-4.65 (m, 2H), 3.86 (d, J=7.5 Hz,1H), 3.57-3.49 (m, 1H), 2.68 (m, 1H), 2.16-1.86 (m, 6H), 1.81-1.77 (m,2H). MS: 645.2 (M+1)⁺.

(2S,4R)—N—((S)-1-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 127

¹H NMR (400 MHz, CDCl₃): δ 8.99 (d, J=4.8 Hz, 1H), 7.62 (d, J=8.7 Hz,1H), 7.49-7.35 (m, 2H), 7.22 (td, J=7.8, 1.5 Hz, 1H), 7.07 (t, J=7.1 Hz,1H), 6.98 (dd, J=7.8, 1.3 Hz, 1H), 6.91 (d, J=8.2 Hz, 1H), 6.72 (tt,J=8.6, 2.2 Hz, 1H), 6.48 (s, 1H), 5.64 (d, J=7.7 Hz, 1H), 4.94-4.69 (m,2H), 4.11-3.91 (m, 1H), 3.46 (s, 1H), 2.79 (m, 1H), 2.19-1.85 (m, 7H),1.61-1.40 (m, 2H). MS: 645.2 (M+1)⁺.

(2S,4R)—N-(1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-cyano-5-fluorophenyl)-1-(4-cyanopyridin-2-yl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 169

¹H NMR (400 MHz, CDCl₃): δ 8.87-8.72 (m, 1H), 8.67-8.48 (m, 1H),8.26-8.01 (m, 1H), 7.56-7.30 (m, 4H), 7.27-7.17 (m, 1H), 7.10 (m, 1H),6.95 (t, J=7.3 Hz, 1H), 6.52-6.28 (m, 1H), 6.21-5.95 (m, 1H), 4.88-4.64(m, 2H), 4.30 (m, 1H), 3.21-2.81 (m, 3H), 2.74-2.19 (m, 3H), 2.13-1.91(m, 1H). MS: 623.1 (M+1)⁺.

(2S,4S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyano-pyridin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 118

¹H NMR (400 MHz, CD₃OD): δ 8.97 (d, J=4.7 Hz, 1H), 7.81-7.62 (m, 2H),7.41-7.35 (m, 2H), 7.26-6.96 (m, 5H), 6.46 (d, J=12.0 Hz, 1H), 4.81-4.75(m, 1H), 4.37-4.28 (m, 1H), 4.25-4.15 (m, 1H), 2.91 (s, 2H), 2.60-2.37(m, 3H), 2.00-1.87 (m, 1H). MS: 598.1 (M+1)⁺.

(2S,4S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyano-pyrimidin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 119

¹H NMR (400 MHz, CD₃OD): δ 8.97 (d, J=4.7 Hz, 1H), 7.81-7.62 (m, 2H),7.41-7.35 (m, 2H), 7.26-6.96 (m, 5H), 6.46 (d, J=12.0 Hz, 1H), 4.81-4.75(m, 1H), 4.37-4.28 (m, 1H), 4.25-4.15 (m, 1H), 2.91 (s, 2H), 2.60-2.37(m, 3H), 2.00-1.87 (m, 1H). MS: 599.1 (M+1)⁺.

(2S,4S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyano-pyridin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 172

¹H NMR (400 MHz, CDCl₃): δ 8.87-8.57 (m, 2H), 7.96 (s, 1H), 7.50-7.30(m, 3H), 7.26-7.12 (m, 2H), 7.09-6.96 (m, 2H), 6.28 (s, 1H), 5.67 (d,J=7.6 Hz, 1H), 4.74 (dd, J=8.1, 4.6 Hz, 1H), 4.42-4.36 (m, 1H), 4.04 (s,1H), 3.87 (d, J=7.8 Hz, 1H), 2.54-2.41 (m, 1H), 2.22-1.76 (m, 8H),1.50-1.32 (m, 2H). MS: 626.2 (M+1)⁺.

(2S,4S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyrimidin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 189

¹H NMR (400 MHz, CDCl₃): δ 9.00 (d, J=4.7 Hz, 1H), 7.76 (s, 1H),7.47-7.30 (m, 2H), 7.24-6.88 (m, 6H), 6.47 (d, J=6.7 Hz, 1H), 5.54 (s,1H), 4.74 (s, 1H), 4.35 (s, 1H), 3.99 (s, 1H), 3.72 (d, J=34.8 Hz, 1H),2.58-2.18 (m, 2H), 1.88 (m, 4H), 1.56-1.42 (m, 2H). MS: 627.2 (M+1)⁺.

(2S,4S)—N—((S)-1-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyano-pyridin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 171

¹H NMR (400 MHz, CDCl₃): δ 8.68 (s, 1H), 8.52 (d, J=5.0 Hz, 1H), 7.60(d, J=8.2 Hz, 1H), 7.45-7.17 (m, 4H), 7.15-6.91 (m, 2H), 6.84 (d, J=8.7Hz, 1H), 6.69 (t, J=8.7 Hz, 1H), 6.54-6.36 (m, 2H), 4.87-4.60 (m, 1H),4.31 (m, 2H), 3.99-3.77 (m, 1H), 3.15-2.78 (m, 2H), 2.62-2.26 (m, 3H),2.26-2.08 (m, 1H). MS: 616.1 (M+1)⁺.

(2S,4S)—N—((S)-1-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 174

¹H NMR (400 MHz, CDCl₃): δ 8.75 (s, 1H), 8.53 (d, J=4.5 Hz, 1H), 7.62(s, 1H), 7.44-7.18 (m, 3H), 7.09-6.96 (m, 2H), 6.86 (s, 1H), 6.71 (t,J=8.7 Hz, 1H), 6.38 (s, 1H), 5.58 (d, J=7.6 Hz, 1H), 4.80 (dd, J=8.0,5.2 Hz, 1H), 4.37 (d, J=5.6 Hz, 1H), 3.96 (s, 1H), 3.61 (d, J=7.7 Hz,1H), 2.62-2.29 (m, 1H), 2.13 (m, 6H), 1.48 (m, 2H). MS: 644.2 (M+1)⁺.

Example 19 Preparation of(2S)—N—((R)-1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-4-methyl-5-oxopyrrolidine-2-carboxamide(Single Enantiomer)—Compound 183

Step A. (2S)-1-tert-Butyl 2-methyl4-(tert-butyldimethylsilyloxy)-4-methyl-5-oxopyrrolidine-1,2-dicarboxylate

LiHMDS (1 M in THF, 22.6 mL, 22.6 mmol) was added into a mixture of(2S,4R)-1-tert-butyl 2-methyl4-(tert-butyldimethylsilyloxy)-5-oxopyrrolidine-1,2-dicarboxylate (6.5g, 17.4 mmol) in THF (60 mL) at −78° C. under N₂. The mixture wasstirred at −78° C. for 1 hr. A solution of iodomethane (2.7 g, 19.1mmol) in THF (10 mL) was added dropwise to the above mixture over 30min. Then the solution was stirred at −78° C. for another 25 min. Theresulting mixture was allowed to warm to room temperature and stirredovernight. The mixture was quenched with NH₄Cl and extracted by ethylacetate (60 mL×3). The combined organic layers were dried andconcentrated. The residue was purified by column chromatography to givethe desired product. MS: 388 (M+1)⁺.

Step B. (2S,4S)-Methyl4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-2-carboxylate

A solution of (2S)-1-tert-butyl 2-methyl4-(tert-butyldimethylsilyloxy)-4-methyl-5-oxopyrrolidine-1,2-dicarboxylate(960 mg, 25 mmol) in TFA/DCM (V:V=1:3) was stirred at room temperaturefor 1 h. The mixture was then concentrated to give the desired productwhich was used directly in the next step. MS: 288 (M+1)⁺.

Step C.(2S)-4-(tert-Butyldimethylsilyloxy)-4-methyl-5-oxopyrrolidine-2-carboxylicacid

To a solution of (2S)-methyl4-(tert-butyldimethylsilyloxy)-4-methyl-5-oxopyrrolidine-2-carboxylate(400 mg, 1.4 mmol) in MeOH/THF/H₂O (V:V:V=2:2:1) was added LiOH (50 mg,2.1 mmol). The mixture was stirred at room temperature for 1 hr and thenconcentrated. The residue was partitioned between ethyl acetate andwater. The aqueous phase was separated and adjusted to pH=3-4 with 1NHCl solution. The aqueous layer was then extracted with ethyl acetate(2×20 mL). The combined organic layers were dried over anhydrous Na₂SO₄and concentrated to give the desired product. MS: 274 (M+1)⁺.

Step D.(2S)-4-(tert-Butyldimethylsilyloxy)-N-(1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-4-methyl-5-oxopyrrolidine-2-carboxamide

A solution of 3-fluoroaniline (83 mg, 0.75 mmol) and2-chlorobenzaldehyde (105 mg, 0.75 mmol) in MeOH (5 mL) was stirred for30 min at room temperature, followed by addition of(2S)-4-(tert-butyldimethylsilyloxy)-4-methyl-5-oxopyrrolidine-2-carboxylicacid (205 mg, 0.75 mmol). The resulting mixture was stirred for 10 minand followed by the addition of 1,1-difluoro-3-isocyanocyclobutane (105mg, 0.9 mmol). The mixture was stirred at room temperature overnight andconcentrated, and then the residue was purified by a standard method togive the desired product. MS: 624 (M+1)⁺.

Step E.(2S)-4-(tert-Butyldimethylsilyloxy)-N-(1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-4-methyl-5-oxopyrrolidine-2-carboxamide

A mixture consisting of(2S)-4-(tert-butyldimethylsilyloxy)-N-(1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-4-methyl-5-oxopyrrolidine-2-carboxamide(200 mg, 0.32 mmol), 2-bromoisonicotinonitrile (88 mg, 0.48 mmol),Cs₂CO₃ (146 mg, 0.45 mmol), Pd₂(dba)₃ (29 mg, 0.032 mmol), Xant-Phos (19mg, 0.032 mmol) and 1,4-dioxane (5 mL) was stirred under N₂ at 80° C.overnight. After filtration, the filtrate was concentrated in vacuo andthe residue was purified by a standard method to give desired product.MS: 726 (M+1)⁺.

Step F.(2S)—N—((R)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-4-methyl-5-oxopyrrolidine-2-carboxamide

To a solution of(2S)-4-(tert-butyldimethylsilyloxy)-N-(1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-4-methyl-5-oxopyrrolidine-2-arboxamide(50 mg, 0.07 mmol) in THF (2 mL) was added TBAF (36 mg, 0.14 mmol) at 0°C. The solution was stirred at 0° C. for 30 min and then partitionedbetween water and EtOAc. Combined organic layers were separated, dried,and concentrated in vacuo. The resulting residue was purified by astandard method to give the desired product. ¹H NMR (400 MHz, CDCl₃): δ8.57 (d, J=5.0 Hz, 1H), 8.48 (d, J=3.8 Hz, 1H), 7.54-7.17 (m, 5H),6.98-6.84 (m, 3H), 6.67 (dd, J=8.6 Hz, 1H), 6.33 (d, J=5.2 Hz, 1H),6.08-6.01 (m, 1H), 4.55-4.48 (m, 1H), 4.29 (s, 1H), 3.22-2.35 (m, 6H),1.93-1.80 (m, 1H), 1.27 (s, 3H). MS: 612.2 (M+1)⁺.

Example 20 Preparation of(2S)—N-(1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxo-ethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluoro-5-sulfamoylphenyl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 158

Step A. Benzyl(3-fluoro-5-nitrophenyl)sulfane

To a solution of 1,3-difluoro-5-nitrobenzene (15.9 g, 100 mmol) in DMF(160 mL) was added K₂CO₃ (15.8 g, 110 mmol) and phenylmethanethiol (12.4g, 100 mmol) at 0° C. The reaction was stirred at room temperature for 2hr and then quenched with H₂O. The resulting mixture extracted withEtOAc (3×100 mL). The combined organic layers were dried over anhydrousNa₂SO₄ and concentrated in vacuo to afford the crude product as a yellowoil which was used in the next step without further purification.

Step B. 3-Fluoro-5-nitrobenzene-1-sulfonyl chloride

To a solution of benzyl(3-fluoro-5-nitrophenyl)sulfane (3.0 g) in DCM(30 mL) was added deionized water (30 mL). Then chlorine was bubbledslowly into the mixture until the complete consumption of the startingmaterial was observed (monitored by TLC). The organic layer wasseparated, washed with sat. aq. Na₂S₂O₃ solution, dried and concentratedto afford the crude product which was used in the next step withoutfurther purification.

Step C. N-tert-butyl-3-fluoro-5-nitrobenzenesulfonamide

To a solution of 3-fluoro-5-nitrobenzene-1-sulfonyl chloride in drydioxane (30 mL) was slowly added tert-butylamine (10 mL) at 0° C. Thereaction was allowed to warm to room temperature and stirred for 2 hr.The mixture was then concentrated and the residue was purified by columnchromatography to afford the desired product. ¹H NMR (400 MHz, DMSO-d₆):δ 8.43 (s, 1H), 8.40-8.32 (m, 1H), 8.10-8.05 (m, 1H), 7.99 (s, 1H), 1.12(s, 9H).

Step D. 3-Amino-N-tert-butyl-5-fluorobenzenesulfonamide

N-tert-butyl-3-fluoro-5-nitro-benzenesulfonamide (1.0 g, 3.6 mmol), ironpowder (1.0 g, 18 mmol) and NH₄Cl (1.0 g, 18 mmol) were mixed in EtOH(95%, 10 mL). The mixture was refluxed for 16 hr then filtered. Thefiltrate was concentrated and the residue was purified by a standardmethod to afford the desired product. ¹H NMR (400 MHz, DMSO-d₆): δ 7.45(s, 1H), 6.88-6.85 (m, 1H), 6.66-6.62 (m, 1H), 6.48-6.42 (m, 1H), 5.89(s, 2H), 1.11 (s, 9H).

Step E

The same as general procedures for UGI reaction set forth herein.

Step F

The same as general procedures for Buchwald reaction set forth herein.

Step G.(S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(3-fluoro-5-sulfamoylphenyl)-5-oxopyrrolidine-2-carboxamide

To a solution of(2S)—N-(3-(N-tert-butylsulfamoyl)-5-fluorophenyl)-N-(1-(2-chlorophenyl)-2-(3,3-difluorocyclo-butylamino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide(80 mg, 0.11 mmol) in DCM (1 mL) was added TFA (1 mL). The reaction wasstirred at room temperature for 16 hr and neutralized with saturated aq.NaHCO₃. The mixture was then extracted with EtOAc (3×10 mL). Thecombined organic layers were dried and concentrated. The residue waspurified by a standard method to afford the target compound. ¹H NMR (400MHz, DMSO-d₆): δ 8.90-8.84 (m, 1H), 8.67-8.62 (m, 1H), 8.55 (s, 1H),8.19 (s, 1H), 7.87-7.76 (m, 1H), 7.65-7.60 (m, 2H), 7.45-7.40 (m, 3H),7.21 (d, J=7.0 Hz, 2H), 7.11-7.04 (m, 1H), 6.93-6.86 (m, 1H), 6.33-6.26(m, 1H), 4.83 (m, 1H), 4.13 (s, 1H), 2.94 (m, 2H), 2.63-2.53 (m, 3H),2.42-2.32 (m, 1H), 1.97 (s, 2H). MS: 661 (M+1)⁺.

Example 21. Preparation of(2S)—N-(1H-benzo[d]imidazol-7-yl)-N-(1-(2-chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide(Racemic)—Compound 141

Step A: 7-Nitro-1H-benzo[d]imidazole

A solution of 3-nitrobenzene-1,2-diamine (900 mg, 5.88 mmol) in AcOH (12mL) was stirred at 100° C. overnight. The mixture was neutralized withaq. NaHCO₃ to pH=8 at 0° C. and the precipitate was collected byfiltration. The precipitate was dried in vacuo to afford the desiredproduct.

Step B:7-Nitro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole NaH(331 mg, 8.28 mmol) was added to a solution of7-nitro-1H-benzo[d]imidazole (900 mg, 5.52 mmol) in DMF (7 mL) at 0° C.under N₂. After stirring at 0° C. for 1 hr, SEMCl (1.38 g, 8.28 mmol)was added and the resulting mixture was stirred at room temperature for2 hr. The reaction mixture was quenched with H₂O and extracted withEtOAc (3×30 mL). The combined organic layers were washed with brine,dried over anhydrous Na₂SO₄, and concentrated. The residue was purifiedby column chromatography to afford the desired product as a yellow oilStep C: 1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-7-amine

To a solution of7-nitro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole (600mg, 2.05 mmol) in EtOH/EtOAc (10 mL/2 mL) was added Pd/C (60 mg). Afterstifling under a hydrogen atmosphere at room temperature overnight, thereaction mixture was filtered and the filtrate was concentrated. Theresidue was purified by a standard method to afford the desired product.

Step D

The same as general procedure for UGI reaction set forth herein.

Step E:(2S)—N-(1H-Benzo[d]imidazol-7-yl)-N-(1-(2-chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide

TBAF (1 M in THF, 3 mL) was added to a solution of(2S)—N-(1-(2-chlorophenyl)-2-((4,4-difluoro-cyclohexyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-5-oxo-N-(1-((2-(trimethylsilyl)ethoxy)-methyl)-1H-benzo[d]imidazol-7-yl)pyrrolidine-2-carboxamidein THF (0.5 mL) at 0° C. under N₂. After stirring at room temperaturefor 7 hr, the reaction was quenched with water at 0° C. The resultingmixture was extracted with EtOAc (3×10 mL). The combined organic layerswere washed with brine, dried over anhydrous Na₂SO₄ and concentrated.The resulting residue was purified by a standard method to afford thedesired product. ¹H NMR (400 MHz, CDCl₃): δ 13.08 (s, 1H), 8.92-8.39 (m,2H), 8.19 (m, 1H), 7.82 (m, 1H), 7.51-7.31 (m, 2H), 7.25 (d, J=5.2 Hz,1H), 7.13-6.70 (m, 3H), 6.41 (m, 1H), 6.20-5.29 (m, 1H), 4.85 (m, 1H),3.86 (s, 1H), 2.97-2.39 (m, 2H), 2.36-1.70 (m, 9H), 1.40 (m, 2H). MS:632.2 (M+1)⁺.

Example 22. Preparation of(4S)—N-(1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-1-methyl-2-oxoimidazolidine-4-carboxamide(Racemic)—Compound 79

Step A: (S)-3-(Benzyloxycarbonyl)-2-oxoimidazolidine-4-carboxylic acid

To a solution of 6.6 g of sodium hydroxide in 140 mL of water at 0° C.,8.8 g of bromine was added dropwise, followed by addition of(S)-4-amino-2-(benzyloxycarbonylamino)-4-oxobutanoic acid (13.4 g, 50mmol) portionwise over 3 min. The resulting yellow solution was heatedto 50° C. for 1 hr and then cooled to room temperature. After additionof sodium thiosulfate (2.0 g), the reaction mixture was washed withether (2×30 mL). The aqueous layer was acidified to pH 1-2 with 6 N HCl.After the precipitate was formed, the suspension was filtered. Thesticky material was collected and re-crystallized in MeOH to afford thedesired product as a white solid. ¹H NMR (400 MHz, DMSO-d₆): δ 13.29 (s,1H), 7.57 (s, 1H), 7.40-7.27 (m, 4H), 5.27-5.04 (m, 2H), 4.66 (dd,J=10.2, 3.2 Hz, 1H), 3.63 (t, J=10.0 Hz, 1H), 3.20 (dd, J=9.7, 3.2 Hz,1H).

Step B: (S)-Dibenzyl 2-oxoimidazolidine-1,5-dicarboxylate

To a 500 mL-flask were added(S)-3-(benzyloxycarbonyl)-2-oxoimidazolidine-4-carboxylic acid (5.3 g,20 mmol), BnBr (2.8 mL, 23 mmol), K₂CO₃ (8.28 g, 60 mmol), andacetonitrile (250 mL). The reaction solution was heated to reflux for 6hr, cooled and then filtered. The filtrate was concentrated in vacuo andthe residue was purified by column chromatography to afford the desiredproduct as white solid. ¹H NMR (400 MHz, CDCl₃): δ 7.41-7.25 (m, 10H),6.36 (s, 1H), 5.30-5.05 (m, 4H), 4.80 (dd, J=10.2, 3.6 Hz, 1H), 3.74 (t,J=10.0 Hz, 1H), 3.41 (dd, J=9.7, 3.7 Hz, 1H).

Step C: (S)-Dibenzyl 3-methyl-2-oxoimidazolidine-1,5-dicarboxylate

To a dry 100 mL-flask were added (S)-dibenzyl2-oxoimidazolidine-1,5-dicarboxylate (1.5 g, 4.24 mmol), K₂CO₃ (1.17 g,8.47 mmol), MeI (5.2 mL, 84.7 mmol), and acetone (50 mL). The reactionsolution was heated to reflux and stirred overnight. The resultingreaction mixture was cooled and filtered. The filtrate was concentratedin vacuo and the residue was purified by column chromatography to affordthe desired product as a white solid. ¹H NMR (400 MHz, CDCl₃): δ7.40-7.26 (m, 10H), 5.27-5.07 (m, 4H), 4.70 (dd, J=10.2, 3.8 Hz, 1H),3.63 (dd, J=10.1, 9.7 Hz, 1H), 3.31 (dd, J=9.6, 3.8 Hz, 1H), 2.84 (s,3H). MS: 369 (M+1)⁺.

Step D: (S)-1-Methyl-2-oxoimidazolidine-4-carboxylic acid

To a dry 50 mL-flask were added (S)-dibenzyl2-oxoimidazolidine-1,5-dicarboxylate (0.5 g, 1.36 mmol), Pd/C (10%, 100mg) and MeOH (15 mL). The suspension was stirred overnight at roomtemperature under a hydrogen atmosphere. The resulting reaction mixturewas filtered. The filtrate was concentrated in vacuo to afford thedesired product as an off-white solid. ¹H NMR (400 MHz, CD₃OD): δ 4.21(dd, J=9.9, 4.8 Hz, 1H), 3.70 (t, J=9.6 Hz, 1H), 3.55 (dd, J=9.3, 4.8Hz, 1H), 2.74 (s, 3H). MS: 145 (M+1)⁺.

Step E:(4S)—N-(1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluoro-phenyl)-1-methyl-2-oxoimidazolidine-4-carboxamide

A mixture of 2-chlorobenzaldehyde (165 mg, 1.18 mmol) and3-fluorobenzenamine (131 mg, 1.18 mmol) in MeOH (3 mL) was stirred atroom temperature for 30 min. Then(S)-1-methyl-2-oxoimidazolidine-4-carboxylic acid (170 mg, 1.18 mmol)was added and the reaction mixture was stirred for another 15 min,followed by addition of 1,1-difluoro-3-isocyanocyclobutane (138 mg, 1.18mmol). The reaction mixture was stirred overnight and concentrated invacuo. The residue was purified by a standard method to give the desiredproduct. MS: 495 (M+1)⁺.

Step F

The same as the Buchwald reaction procedure set forth herein. ¹H NMR(400 MHz, CDCl₃): δ 8.64-8.34 (m, 2H), 7.94-7.59 (m, 1H), 7.50-6.61 (m,8H), 6.34-6.07 (m, 1H), 4.94-4.67 (m, 1H), 4.3-4.2 (m, 1H), 3.49 (m,1H), 3.46-3.22 (m, 1H), 3.02-2.83 (m, 2H), 2.87 (s, 3H), 2.5-2.2 (m,2H). MS: 597 (M+1)⁺.

Example 23. Preparation of(S)—N—((S)-1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-2-oxoimidazolidine-4-carboxamide(Single Enantiomer)—Compound 80

Step A: (S)-3,4-Dibenzyl 1-tert-butyl2-oxoimidazolidine-1,3,4-tricarboxylate

To a 25 mL-flask were added (S)-dibenzyl2-oxoimidazolidine-1,5-dicarboxylate (40 mg, 0.11 mmol), (BOC)₂O (26 mg,0.12 mmol), TEtOAc (0.06 mL, 0.3 mmol), DMAP (cat.) and CH₂Cl₂ (2 mL).The mixture was stirred overnight. The solvent was then removed in vacuoand the residue was purified by column chromatography to give thedesired product. ¹H NMR (400 MHz, CDCl₃): δ 7.39-7.27 (m, 10H), 5.24 (s,2H), 5.16 (s, 2H), 4.67 (dd, J=10.2, 3.5 Hz, 1H), 3.94 (dd, J=11.1, 10.3Hz, 1H), 3.74 (dd, J=11.2, 3.5 Hz, 1H), 1.51 (s, 9H).

Step B: (S)-1-(tert-Butoxycarbonyl)-2-oxoimidazolidine-4-carboxylic acid

To a dry 50 mL-flask were added (S)-3,4-dibenzyl 1-tert-butyl2-oxoimidazolidine-1,3,4-tricarboxylate (1.24 g, 2.73 mmol), Pd/C (10%,200 mg) and MeOH (30 mL). The suspension was stirred overnight at roomtemperature under a hydrogen atmosphere. The reaction mixture wasfiltered, and the filtrate was concentrated in vacuo to afford thedesired product. ¹H NMR (400 MHz, DMSO-d₆): δ 6.06 (s, 2H), 4.31 (s,1H), 4.25-3.94 (m, 2H), 1.52 (s, 9H).

Step C: (4S)-tert-Butyl4-((1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)(3-fluorophenyl)carbamoyl)-2-oxoimidazolidine-1-carboxylate

A mixture of 2-chlorobenzaldehyde (122 mg, 0.87 mmol) and3-fluorobenzenamine (97 mg, 0.87 mmol) in MeOH (2 mL) was stirred atroom temperature for 30 min. Then(S)-1-(tert-butoxycarbonyl)-2-oxoimidazolidine-4-carboxylic acid (200mg, 0.87 mmol) was added and the reaction mixture was stirred foranother 15 min followed by addition of1,1-difluoro-3-isocyanocyclobutane (102 mg, 0.87 mmol). The reactionmixture was further stirred at room temperature overnight and thenconcentrated in vacuo. The residue was purified by a standard method togive the desired product. ¹H NMR (400 MHz, CDCl₃): δ 7.46-6.59 (m, 8H),6.45 (s, 1H), 4.41-4.04 (m, 2H), 4.01-3.78 (m, 1H), 3.64-3.30 (m, 1H),2.92 (m, 2H), 2.71-2.27 (m, 2H), 1.46 (s, 9H). MS: 581 (M+1)⁺.

Step D: (4S)-tert-Butyl4-((1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)(3-fluorophenyl)carbamoyl)-3-(4-cyanopyridin-2-yl)-2-oxoimidazolidine-1-carboxylate

To a 25 mL flask charged with 1,4-dioxane (4.5 mL) were added(4S)-tert-butyl 4-((1-(2-chlorophenyl)-2-(3,3-difluoro cyclobutylamino)-2-oxoethyl)(3-fluoro-phenyl)carbamoyl)-2-oxoimidazolidine-1-carboxylate(250 mg, 0.43 mmol), 2-bromoisonicotinonitrile (122 mg, 0.65 mmol),Cs₂CO₃ (281 mg, 0.862 mmol), Xant-Phos (25 mg, 0.043 mmol) and Pd₂(dba)₃(40 mg, 0.043 mmol). The mixture was degassed and refilled withnitrogen, and then heated to 100° C. for 3 hr. The resulting mixture wascooled and filtered. The filtrate was concentrated in vacuo and theresidue was purified by a standard method to give both epimers. Theepimers were further separated by a standard method to give the desiredproduct. ¹H NMR (400 MHz, CDCl₃): δ 8.58 (s, 1H), 8.48 (t, J=5.9 Hz,1H), 7.71 (d, J=8.4 Hz, 1H), 7.37-7.16 (m, 4H), 7.15-6.76 (m, 4H),6.56-6.31 (m, 2H), 4.95-4.75 (m, 1H), 4.31 (s, 1H), 3.86 (dd, J=10.8,5.1 Hz, 1H), 3.66 (m, 1H), 2.99 (m, 2H), 2.61-2.27 (m, 2H), 1.56 (s,9H). MS: 683 (M+1)⁺.

Step E: (S)—N—((S)-1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-2-oxoimidazolidine-4-carboxamide

To a solution of 2N HCl/MeOH (2 mL) at 0° C. was added 50 mg of(S)-tert-butyl-4-(((S)-1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)(3-fluorophenyl)carbamoyl)-3-(4-cyanopyridin-2-yl)-2-oxoimidazolidine-1-carboxylate. Thereaction mixture was warmed to room temperature and stirred for 5 hr.The solvent was removed in vacuo and the residue was purified by astandard method to give the desired product. ¹H NMR (400 MHz, CD₃OD): δ8.50 (d, J=4.5 Hz, 1H), 7.65 (d, J=8.6 Hz, 1H), 7.50-6.81 (m, 8H), 6.47(d, J=11.6 Hz, 1H), 5.04-4.92 (m, 1H), 4.22 (m, 1H), 3.59-3.46 (m, 1H),3.39 (dd, J=9.9, 4.5 Hz, 1H), 2.91 (m, 2H), 2.63-2.36 (m, 2H). MS: 583(M+1)⁺.

Example 24. Preparation of(4S)—N-(1-(2-chlorophenyl)-2-(3,3-difluorocyclobutyl-amino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-1-(2-hydroxyethyl)-2-oxoimidazolidine-4-carboxamide(Racemic)

Step A: (S)-Dibenzyl3-(2-ethoxy-2-oxoethyl)-2-oxoimidazolidine-1,5-dicarboxylate

To a dry 50 mL-flask charged with DME (5 mL) were added (S)-dibenzyl2-oxoimidazolidine-1,5-dicarboxylate (200 mg, 0.56 mmol), K₂CO₃ (156 mg,1.13 mmol), and ethyl 2-bromoacetate (0.13 mL, 1.13 mmol). The mixturewas heated to reflux for 3 hr. The reaction mixture was cooled andfiltered. The filtrate was concentrated in vacuo and the residue waspurified by column chromatography to afford the desired product as acolorless oil. ¹H NMR (400 MHz, CDCl₃): δ 7.45-7.25 (m, 10H), 5.41-5.05(m, 4H), 4.80 (dd, J=10.2, 3.5 Hz, 2H), 4.29-4.08 (m, 3H), 3.90 (dd,J=12.2, 7.2 Hz, 2H), 3.45 (dd, J=9.2, 3.5 Hz, 1H), 1.28 (td, J=7.1, 2.1Hz, 3H).

Step B: (S)-1-(2-Ethoxy-2-oxoethyl)-2-oxoimidazolidine-4-carboxylic acid

To a dry 50 mL-flask were added (S)-dibenzyl3-(2-ethoxy-2-oxoethyl)-2-oxoimidazolidine-1,5-dicarboxylate (170 mg,0.386 mmol), Pd/C (10%, 35 mg) and MeOH (4 mL). The suspension wasstirred at room temperature overnight under a hydrogen atmosphere. Thereaction mixture was filtered, and the filtrate was concentrated invacuo to afford the desired product as an off-white solid. ¹H NMR (400MHz, CD₃OD): δ 4.30 (dd, J=10.0, 4.8 Hz, 1H), 4.20 (q, J=7.1 Hz, 2H),4.05-3.91 (m, 2H), 3.91-3.85 (m, 1H), 3.69 (dd, J=9.0, 4.8 Hz, 1H), 1.29(t, J=7.1 Hz, 3H).

Step C: Ethyl2-((4S)-4-((1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)(3-fluorophenyl)carbamoyl)-2-oxoimidazolidin-1-yl)acetate

A mixture of 2-chlorobenzaldehyde (518 mg, 3.70 mmol) and3-fluorobenzenamine (411 mg, 3.7 mmol) in MeOH (12 mL) was stirred atroom temperature for 30 min. Then(S)-1-(2-ethoxy-2-oxoethyl)-2-oxoimidazolidine-4-carboxylic acid (800mg, 3.7 mmol) was added and the reaction mixture was stirred for another30 min, followed by addition of 1,1-difluoro-3-isocyanocyclobutane (600mg, 3.7 mmol). The reaction mixture was stirred overnight andconcentrated in vacuo. The residue was purified by a standard method togive the desired product. MS: 567: (M+1)⁺.

Step D: Ethyl2-((4S)-4-((1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3-fluorophenyl)carbamoyl)-3-(4-cyanopyridin-2-yl)-2-oxoimidazolidin-1-yl)acetate—Compound94

To a 25 mL-flask were added ethyl2-((4S)-4-((1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)(3-fluoro-phenyl)carbamoyl)-2-oxoimidazolidin-1-yl)acetate (50 mg,0.0882 mmol), 2-bromoisonicotinonitrile (21 mg, 0.115 mmol), Cs₂CO₃ (58mg, 0.176 mmol), Xant-Phos (5.2 mg, 0.009 mmol), Pd₂(dba)₃ (8.2 mg,0.009 mmol) and 1,4-dioxane (1 mL). The mixture was degassed andrefilled with nitrogen, and then heated to 100° C. for 3 hr. Theresulting mixture was cooled and filtered and then the filtrate wasconcentrated in vacuo. The residue was purified by a standard method togive both epimers. ¹H NMR (400 MHz, CDCl₃): δ 8.63-8.57 (S, 1H),8.55-8.38 (m, 1H), 7.63 (s, 1H), 7.46-6.84 (m, 8H), 6.45-6.37 (m, 1H),6.22-5.94 (m, 1H), 5.06-4.77 (m, 1H), 4.43-4.37 (m, 1H), 4.32-4.20 (m,1H), 4.21 (q, J=7.1 Hz, 2H), 3.82-3.46 (m, 3H), 3.12-2.82 (m, 2H),2.66-2.25 (m, 2H), 1.29 (t, J=7.1 Hz, 3H). MS: 669 (M+1)⁺.

Step E:(4S)—N-(1-(2-Chlorophenyl)-2-(3,3-difluorocyclobutyl-amino)-2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-1-(2-hydroxyeth-yl)-2-oxoimidazolidine-4-carboxamide—Compound112

To a solution of ethyl2-((4S)-4-((1-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)(3-fluorophenyl)carbamoyl)-2-oxo-3-(pyrimidin-2-yl)imidazolidin-1-yl)acetate(100 mg, 0.155 mmol) in DME (2 mL) at 0° C. was added LiBH₄ (22 mg) intwo portions. The mixture was stirred for 0.5 hr, then warmed to roomtemperature. The resulting mixture was stirred for another 2 hr andquenched with H₂O (2 mL) at 0° C. The resulting mixture was extractedwith EtOAc (2×10 mL). The combined organic layers were combined, washedwith brine, dried over anhydrous Na₂SO₄, and concentrated in vacuo. Theresidue was purified by a standard method to give the desired product.¹H NMR (400 MHz, CDCl₃): δ 8.62-8.55 (m, 2H), 7.63 (d, J=8.1 Hz, 1H),7.40-6.85 (m, 8H), 6.47-6.2 (m, 2H), 4.90-4.69 (m, 1H), 4.30-4.15 (m,1H), 3.87-3.72 (m, 2H), 3.71-3.19 (m, 5H), 3.08-2.85 (m, 2H), 2.63-2.35(m, 2H). MS: 603 (M+1)⁺.

The following compound was synthesized via the procedure set forthabove, using the appropriate aldehyde, amine, carboxylic acid,isocyanide and halo-substituted aromatic ring or heterocyclic(heteroaromatic) ring using the reagents and solvents set forth above,and purified via standard methods.

Ethyl2-((4S)-4-((1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3-fluorophenyl)carbamoyl)-3-(4-cyanopyrimidin-2-yl)-2-oxoimidazolidin-1-yl)acetate(Racemic)—Compound 111

¹H NMR (400 MHz, CDCl₃): δ 8.90-8.82 (m, 1H), 7.62-7.57 (m, 1H),7.46-6.82 (m, 8H), 6.52-6.48 (m, 1H), 6.15-5.85 (m, 2H), 4.88-4.82 (m,1H), 4.45-4.35 (m, 1H), 4.32-4.13 (m, 2H), 3.86-3.46 (m, 3H), 3.05-2.85(m, 2H), 2.56-2.32 (m, 2H), 1.29 (t, J=7.1 Hz, 3H). MS: 670 (M+1)⁺.

Example A: In Vitro Assays for IDH1m (R132H or R132C) Inhibitors

A test compound is prepared as 10 mM stock in DMSO and diluted to 50×final concentration in DMSO, for a 50 μl reaction mixture. IDH enzymeactivity converting alpha-ketoglutarate to 2-hydroxyglutaric acid ismeasured using a NADPH depletion assay. In the assay the remainingcofactor is measured at the end of the reaction with the addition of acatalytic excess of diaphorase and resazurin, to generate a fluorescentsignal in proportion to the amount of NADPH remaining. IDH1-R132homodimer enzyme is diluted to 0.125 μg/ml in 40 μl of Assay Buffer (150mM NaCl, 20 mM Tris-Cl pH 7.5, 10 mM MgCl₂, 0.05% BSA, 2 mMb-mercaptoethanol); 1 μl of test compound dilution in DMSO is added andthe mixture is incubated for 60 minutes at room temperature. Thereaction is started with the addition of 10 μl of Substrate Mix (20 μlNADPH, 5 mM alpha-ketoglutarate, in Assay Buffer) and the mixture isincubated for 90 minutes at room temperature. The reaction is terminatedwith the addition of 25 μl of Detection Buffer (36 μg/ml diaphorase, 30mM resazurin, in 1× Assay Buffer), and is incubated for 1 minute beforereading on a SpectraMax platereader at Ex544/Em590.

Compounds are assayed for their activity against IDH1 R132C followingthe same assay as above with the following modifications: Assay Bufferis (50 mM potassium phosphate, pH 6.5; 40 mM sodium carbonate, 5 mMMgCl₂, 10% glycerol, 2 mM b-mercaptoethanol, and 0.03% BSA). Theconcentration of NADPH and alpha-ketoglutarate in the Substrate Bufferis 20 μM and 1 mM, respectively.

Representative compounds of formula I set forth in Table 1 were testedin this assay or a similar assay and the results are set forth below inTable 2. As used in Table 2, “A” refers to an inhibitory activityagainst IDH1 R132H or IDH1 R132C with an IC₅₀<0.1 μM; “B” refers to aninhibitory activity against IDH1 R132H or IDH1 R132C with an IC₅₀between 0.1 μM and 0.5 μM; “C” refers to an inhibitory activity againstIDH1 R132H or IDH1 R132C with an IC₅₀ between 0.5 μM and 1 μM; “D”refers to an inhibitory activity against IDH1 R132H or IDH1 R132C withan IC₅₀ between 1 μM and 2 μM.

TABLE 2 Inhibitory Activities of Representative Compounds of formula IIDH IDH R132C R132H HT1080 U87MG Cpd IC50 IC50 IC50 IC50 No (uM) (uM)(uM) (uM) 1 A A A B 2 D B 3 B B B 4 A A A A 5 A A A B 6 A B B 7 A A A A8 B C 9 A A A A 10 B B 11 B B 12 A B B 13 C C 14 A A A B 15 A A B B 16 BB B C 17 B B C D 18 A A A A 19 B C 20 A A B B 21 A A A B 22 B B 23 A B BB 24 C D 25 B C 26 A B B 27 A A 28 A B A 29 A A A 30 A A B 31 A B C 32 BD 33 A A A B 34 A B C 35 A B B 36 B B 37 A A A A 38 C D 39 C D 40 A A BB 41 A B C 42 B C 43 A A A A 44 B B 45 A A B B 46 C D 47 A A A B 48 A AB B 49 A A B B 50 C D 51 A B B B 52 A A 53 A A A A 54 B B 55 A A A A 56A A 57 B C 58 A A A A 59 B C 60 B B 61 B B 62 A B 63 A A A A 64 A A A A68 A A A A 69 A A A A 70 A A A A 71 A A A A 72 A A A A 73 A A A A 74 A AA A 75 A A A A 76 A A A A 77 A A A A 78 A A A A 79 A A A A 80 A A A A 81A A A A 82 A A A A 83 A A A A 84 A A A B 85 A A A A 86 A A A A 87 A A AA 88 A A A A 89 A A A A 90 A A A A 91 A A A A 92 A A A A 93 A A A A 94 AA A A 95 A A A A 96 A A A A 97 A A A A 98 A A A A 99 A A A A 100 A A A A101 A A A A 102 A A A B 103 A A A B 104 A A A A 105 A A A A 106 A A A A107 A A A A 108 A A A A 109 A A A B 110 A A A A 111 A A A A 112 A A A A113 A A A A 114 A A A A 115 A A A A 116 A A A B 117 A A A A 118 A A A A119 A A A B 120 A A A B 121 A A A A 122 A A A B 123 A A A A 124 A A A A125 A A A A 126 A A A B 127 A A A A 128 A A A A 129 A A A A 130 A A A A131 A A A A 132 A A A A 133 A A A A 134 A A A A 135 A A A A 136 A A A A137 A A A A 138 A A A A 139 A A A A 140 A A A A 141 A A A A 142 A A A A143 A A A A 144 A A A A 145 A A A A 146 A A A A 147 A A A A 148 A A A A149 A A A A 150 A A A A 151 A A A A 152 A A A A 153 A A A A 154 A A A A155 A A A A 156 A A A A 157 A A A A 158 A A A A 159 A A A A 160 A A A A161 A A A A 162 A A A A 163 A A A A 164 A A A A 165 A A A 166 A A A A167 A A A 168 A A A A 169 A A A 170 A A A 171 A A A 172 A A A 173 A A A174 A A A A 175 A A A A 176 A A A A 177 A A A A 178 A A B A 179 A A A A180 A A A A 181 A A A A 182 A A A A 183 A A A 184 A A A A 185 A A A A186 A A A A 187 A A A A 188 A A A A 189 A A A 190 A A A A 191 A A A A192 A A A 193 A A A A 194 A A A 195 A A A 196 A A A 197 A A A A 198 A AA A 199 A A A A 200 A A A A 201 A A A A 202 A A A 203 A A A 204 A A A A205 A A A A 206 A A A 207 A A A A 208 A A A A 209 A A A A 210 A A A A211 A A A A 212 A A A A

Example B: Cellular Assays for IDH1m (R132H or R132C) Inhibitors

Cells (HT1080 or U87MG) are grown in T125 flasks in DMEM containing 10%FBS, 1× penicillin/streptomycin and 500 ug/mL G418 (present in U87MGcells only). They are harvested by trypsin and seeded into 96 well whitebottom plates at a density of 5000 cell/well in 100 ul/well in DMEM with10% FBS. No cells are placed in columns 1 and 12. Cells are incubatedovernight at 37° C. in 5% CO₂. The next day test compounds are made upat 2× the final concentration and 100 ul are added to each cell well.The final concentration of DMSO is 0.2% and the DMSO control wells areplated in row G. The plates are then placed in the incubator for 48hours. At 48 hours, 100 ul of media is removed from each well andanalyzed by LC-MS for 2-HG concentrations. The cell plate is placed backin the incubator for another 24 hours. At 72 hours post compoundaddition, 10 mL/plate of Promega Cell Titer Glo reagent is thawed andmixed. The cell plate is removed from the incubator and allowed toequilibrate to room temperature. Then 100 ul of Promega Cell Titer Gloreagent is added to each well of media. The cell plate is then placed onan orbital shaker for 10 minutes and then allowed to sit at roomtemperature for 20 minutes. The plate is then read for luminescence withan integration time of 500 ms.

The IC₅₀ for inhibition of 2-HG production (concentration of testcompound to reduce 2HG production by 50% compared to control) in thesetwo cell lines for various compounds of formula I is set forth in Table2 above. As used in Table 2, “A” refers to an IC₅₀ for inhibition of2-HG production ≤0.1 μM; “B” refers to an IC₅₀ for inhibition of 2-HGproduction between 0.1 μM and 0.5 μM; “C” refers to an IC₅₀ forinhibition of 2-HG production between 0.5 μM and 1 μM; “D” refers to anIC₅₀ for inhibition of 2-HG production between 1 μM and 2 μM.

Example C: Metabolic Stabilities of Compounds of Formula I

Metabolic stabilities of compounds of formula I can be tested with thefollowing assay and species specific liver microsomes (LM) extractionratio (Eh) can be calculated:

-   1. Buffer A: 1.0 L of 0.1 M monobasic Potassium Phosphate buffer    containing 1.0 mM EDTA; Buffer B: 1.0 L of 0.1 M Dibasic Potassium    Phosphate buffer containing 1.0 mM EDTA; Buffer C: 0.1 M Potassium    Phosphate buffer, 1.0 mM EDTA, pH 7.4 by titrating 700 mL of buffer    B with buffer A while monitoring with the pH meter.-   2. Reference compounds (Ketanserin) and test compounds spiking    solution: 500 μM spiking solution: add 10 μL of 10 mM DMSO stock    solution into 190 μL CAN; 1.5 μM spiking solution in microsomes    (0.75 mg/mL): add 1.5 μL of 500 μM spiking solution and 18.75 μL of    20 mg/mL liver microsomes into 479.75 μL of Buffer C.-   3. NADPH stock solution (6 mM) is prepared by dissolving NADPH into    buffer C.-   4. Dispense 30 μL 1.5× compound/liver microsome solution in 96-well    plate and immediately add 135 μL ACN containing IS before adding 15    uL Buffer C to prepare real 0 minute samples.-   5. Add 15 μL of NADPH stock solution (6 mM) to the wells designated    as Time 30, and start timing.-   6. At the end of incubation (0 min), add 135 μL of ACN containing    the internal standard Osalmid) to all the wells (30 min, and 0 min).    Then add 15 μL of NADPH stock solution (6 mM) to the wells    designated as Time 0.-   7. After quenching, centrifuge the reaction mixtures at 3220 g for    10 min.-   8. Transfer 50 μL of the supernatant from each well into a 96-well    sample plate containing 50 μL of ultra pure water (Millipore) for    LC/MS analysis.

The invention claimed is:
 1. A method of treating cholangiocarcinomacharacterized by the presence of an IDH1 mutation in a patient in needthereof comprising administering to the patient a therapeuticallyeffective amount of(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide:

or a pharmaceutically acceptable salt or hydrate thereof.
 2. The methodof claim 1 wherein the IDH1 mutation is an IDH1 R132H or R132C mutation.